bims-instec 2021-08-01 papers

bims-instec

Biomed News

on Intestinal stem cells and chemoresistance in colon cancer and intestinal regeneration

Issue of 2021–08–01
29 papers selected by
Maria-Virginia Giolito, IRFAC/UMR-S1113 INSERM

POU4F2 Promotes Colorectal Cancer Cell Migration and Invasion via Hedgehog-mediated Epithelial-Mesenchymal Transition.
PARP-Targeted Auger Therapy in p53 Mutant Colon Cancer Xenograft Mouse Models.
PEITC inhibits the invasion and migration of colorectal cancer cells by blocking TGF-β-induced EMT.
Hydroxyphenyl Butanone Induces Cell Cycle Arrest through Inhibition of GSK3β in Colorectal Cancer.
B-Myb accelerates colorectal cancer progression through reciprocal feed-forward transactivation of E2F2.
A novel class of inhibitors that target SRSF10 and promote p53-mediated cytotoxicity on human colorectal cancer cells.
Identification of cis-HOX-HOXC10 axis as a therapeutic target for colorectal tumor-initiating cells without APC mutations.
Tazarotene-induced gene 1 interacts with Polo-like kinase 2 and inhibits cell proliferation in HCT116 colorectal cancer cells.
Insect Gut Regeneration.
Diverse expression patterns of mucin 2 in colorectal cancer indicates its mechanism related to the intestinal mucosal barrier.
IGF2BP2 promotes the progression of colorectal cancer through a YAP-dependent mechanism.
lncRNA-PACER upregulates COX-2 and PGE2 through the NF-κB pathway to promote the proliferation and invasion of colorectal-cancer cells.
Long noncoding RNA MALAT1 sponging miR-26a-5p to modulate Smad1 contributes to colorectal cancer progression by regulating autophagy.
Longitudinal change of genetic variations in cetuximab-treated metastatic colorectal cancer.
Diosgenin exerts anti-tumor effects through inactivation of cAMP/PKA/CREB signaling pathway in colorectal cancer.
CXCL2-CXCR2 axis mediates αV integrin-dependent peritoneal metastasis of colon cancer cells.
Trop-2 induces ADAM10-mediated cleavage of E-cadherin and drives EMT-less metastasis in colon cancer.
Shedding Light on BRAF: Management of Colorectal Cancer in the Era of Personalized Medicine.
Systems-level biomarkers identification and drug repositioning in colorectal cancer.
Expression and prognostic significance of CBX2 in colorectal cancer: database mining for CBX family members in malignancies and vitro analyses.
Cancer stem cells, epithelial-mesenchymal transition, ATP and their roles in drug resistance in cancer.
Mutational profiles of metastatic colorectal cancer treated with FOLFIRI plus cetuximab or bevacizumab before and after secondary resection (AIO KRK 0306; FIRE-3).
Impact of UGT1A1 Genotype on the Efficacy and Safety of Irinotecan-based Chemotherapy in Metastatic Colorectal Cancer.
An IRF1-dependent pathway of TNFα-induced shedding in intestinal epithelial cells.
Tissue-specific reduction in MLH1 expression induces microsatellite instability in intestine of Mlh1+/- mice.
The prognostic and predictive impact of BRAF mutations in deficient mismatch repair/microsatellite instability-high colorectal cancer: systematic review/meta-analysis.
Discovery of small molecule acting as multitarget inhibitor of colorectal cancer by simultaneous blocking of the key COX-2, 5-LOX and PIM-1 kinase enzymes.
Oxymatrine Synergistically Enhances Doxorubicin Anticancer Effects in Colorectal Cancer.
Cell cycle on the crossroad of tumorigenesis and cancer therapy.

Cancer Sci. 2021 Jul 30.

POU4F2 Promotes Colorectal Cancer Cell Migration and Invasion via Hedgehog-mediated Epithelial-Mesenchymal Transition.

Kaibo Guo, Peipei Wang, Leyin Zhang, Yiwen Zhou, Xinyang Dai, Yici Yan, Yuxuan Chen, Harpreet S Wasan, Jieru Yu, Shanming Ruan, Leitao Sun.

  As a POU homeodomain transcription factor, POU4F2 has been implicated in regulating tumorigenic processes in various cancers. However, the role of POU4F2 in colorectal cancer remains unclear. Here, we revealed that POU4F2 functions as a tumor promotor in colorectal cancer. Bioinformatics analysis in specimens from colorectal cancer patients and expression analysis in colorectal cancer cell lines showed that POU4F2 was upregulated at the mRNA and protein levels in colorectal cancer. Depletion of POU4F2 suppressed the metastatic phenotypes of colorectal cancer cells, including cell migration, invasion, and the expression of epithelial-mesenchymal transition markers. Moreover, depletion of POU4F2 decreased the number of lung metastatic nodes in nude mice. Mechanistically, POU4F2 positively regulated the Hedgehog signaling pathway, as inferred from the downregulation of the expression of SHH, PTCH1, SMO, and GLI-1 in vitro and vivo following silencing of POU4F2. Furthermore, the SMO agonist SAG reversed the effects of POU4F2 knockdown in colorectal cancer. Functionally, POU4F2 contributed to the Hedgehog signaling-regulated activation of the epithelial-mesenchymal transition process and promotion of colorectal cancer cell migration and invasion. Collectively, these findings elucidated the role of POU4F2 as a tumor promotor in colorectal cancer through the regulation of Hedgehog signaling-mediated epithelial-mesenchymal transition and suggested that POU4F2 suppression might be a promising therapeutic target in inhibiting colorectal cancer metastasis.

Keywords:  Colorectal Cancer; EMT; Hedgehog Pathway; Migration and Invasion; POU4F2

DOI:  https://doi.org/10.1111/cas.15089
Mol Pharm. 2021 Jul 28.

PARP-Targeted Auger Therapy in p53 Mutant Colon Cancer Xenograft Mouse Models.

Thomas Wilson, Giacomo Pirovano, Gu Xiao, Zachary Samuels, Sheryl Roberts, Tara Viray, Navjot Guru, Pat Zanzonico, Marc Gollub, Naga Vara Kishore Pillarsetty, Thomas Reiner, Jill Bargonetti.

  Despite Auger electrons being highly appealing due to their short-range and high linear energy transfer to surrounding tissues, the progress in the field has been limited due to the challenge in delivering a therapeutic dose within the close proximity of cancer cell's DNA. Here, we demonstrate that the PARP inhibitor 123I-MAPi is a viable agent for the systemic administration and treatment of p53 mutant cancers. Significantly, minimal off-site toxicity was observed in mice administered with up to 74 MBq of 127I-PARPi. Taken together, these results lay the foundation for future clinical evaluation and broader preclinical investigations. By harnessing the scaffold of the PARP inhibitor Olaparib, we were able to deliver therapeutic levels of Auger radiation to the site of human colorectal cancer xenograft tumors after systemic administration. In-depth toxicity studies analyzed blood chemistry levels and markers associated with specific organ toxicity. Finally, p53+/+ and p53-/- human colorectal cancer cell lines were evaluated for the ability of 123I-MAPi to induce tumor growth delay. Toxicity studies demonstrate that both 123I-MAPi and its stable isotopologue, 127I-PARPi, have no significant off-site toxicity when administered systemically. Analysis following 123I-MAPi treatment confirmed its ability to induce DNA damage at the site of xenograft tumors when administered systemically. Finally, we demonstrate that 123I-MAPi generates a therapeutic response in p53-/-, but not p53+/+, subcutaneous xenograft tumors in mouse models. Taken together, these results represent the first example of a PARP Auger theranostic agent capable of delivering a therapeutic dose to xenograft human colorectal cancer tumors upon systemic administration without causing significant toxicity to surrounding mouse organs. Moreover, it suggests that a PARP Auger theranostic can act as a targeted therapeutic for cancers with mutated p53 pathways. This landmark goal paves the way for clinical evaluation of 123I-MAPi for pan cancer therapeutics.

Keywords:  123I-MAPi; Auger; PARP; p21; p53

DOI:  https://doi.org/10.1021/acs.molpharmaceut.1c00323
Biomed Pharmacother. 2020 Oct;pii: S0753-3322(20)30936-7. [Epub ahead of print]130 110743

PEITC inhibits the invasion and migration of colorectal cancer cells by blocking TGF-β-induced EMT.

Jian Xiao, Ningning Zhou, Yin Li, Yunyun Xiao, Wei Chen, Junwen Ye, Tenghui Ma, Yan Zhang.

   AIM: Tumor metastasis is the leading cause of death in patients with colorectal cancer (CRC), in which epithelial-mesenchymal transition(EMT) plays a vital role. However, the exact mechanisms of this process remain largely unknown. The aim of the present study was to determine the role of phenethyl isothiocyanate (PEITC) in CRC metastasis by regulating EMT.
MAIN METHODS: Wound healing assays and Transwell matrix assays were used to evaluate the potential of PEITC to inhibit CRC cells invasion and migration in vitro. Western blotting, light microscopy and immunofluorescence assays were used to detect the occurrence of EMT. Luciferase activity assay, real time-PCR and western blotting were used to investigate TGF-β1/Smad signaling activity.
KEY FINDINGS: We observed that PEITC, an isothiocyanate compound from crucifer with chemopreventive potential, inhibited the invasion and migration of CRC cells. Moreover, we showed that PEITC regulated the EMT of CRC cells. Additionally, we demonstrated that PEITC blocked the activation of the TGF-β1/Smad pathway and significantly suppressed TGF-β1-induced EMT.
SIGNIFICANCE: Our results suggested that PEITC plays a crucial role in inhibiting the invasion and migration of CRC cells by regulating TGF-β1-induced EMT. The results of the present study provide a theoretical basis for the use of PEITC to treat CRC.

Keywords:  Colorectal cancer; EMT; Invasion; Migration; PEITC; TGF-β1/Smad

DOI:  https://doi.org/10.1016/j.biopha.2020.110743
Biomed Res Int. 2021 ;2021 9981815

Hydroxyphenyl Butanone Induces Cell Cycle Arrest through Inhibition of GSK3β in Colorectal Cancer.

Songyan Zhang, Yunfeng Wang, Haopeng Zhang, Chengming Sun, Shuwei Dang, Ming Liu.

Background: Colorectal cancer (CRC) is among the top three gastrointestinal malignancy in morbidity and mortality. The abnormal activation of Wnt/β-catenin pathway is considered to be a key factor in the occurrence and development of CRC. Novel inhibitor discovery against key factor in WNT pathway is important for CRC treatment and prevention.
Methods: Cell proliferation was detected after hydroxyphenyl butanone treatment in human colorectal cancer HCT116, LOVO, and normal colonic epithelial NCM460 cells. Colony formation, cell invasion ability, and cell cycle were detected with and without GSK-3β knockdown.
Results: Hydroxyphenyl butanone induces cycle arresting on G1-S phase of colorectal cancer cell line through GSK3β in Wnt/β-catenin pathway and inhibits malignant biological manifestations of cell proliferation, colony formation, and invasion. The inhibition in the high concentration group is stronger than that in the low concentration group, and the antitumor effect is different for different tumor cells. Under the same concentration of natural hydroxyphenyl butanone, the inhibition on normal colonic epithelial cells is significantly lower than that on tumor cells. The natural hydroxyphenyl butanone with medium and low concentration could promote the proliferation of normal colonic epithelial cells.
Conclusion: This study illustrated natural hydroxyphenyl butanone as new inhibitor of GSK3β and revealed the mechanisms underlying the inhibitory effects in colorectal cancer.

DOI: https://doi.org/10.1155/2021/9981815
Oncogene. 2021 Jul 27.

B-Myb accelerates colorectal cancer progression through reciprocal feed-forward transactivation of E2F2.

Xiaoyan Fan, Yitao Wang, Tinghui Jiang, Tao Liu, Yuelei Jin, Kailong Du, Yulong Niu, Chunxue Zhang, Zhongyu Liu, Yunlong Lei, Youquan Bu.

B-Myb is an important transcription factor that plays a critical role in gene expression regulation and tumorigenesis. However, its functional implication in colorectal cancer remains elusive. In this study, we found that B-Myb was significantly upregulated at both mRNA and protein levels in colorectal cancer samples compared to non-tumor counterparts. B-Myb overexpression accelerated cell proliferation, cell cycle progression and cell motility in colorectal cancer cells, and promoted tumor growth in orthotopic nude mouse models in vivo. In contrast, B-Myb depletion inhibited these malignant phenotypes. Mechanistic investigations revealed that E2F2 was a novel transcriptional target of B-Myb and is essential to B-Myb-induced malignant phenotypes. Notably, B-Myb and E2F2 exhibited positive expression correlation, and interacted with each other in colorectal cancer cells. In addition to their autoregulatory mechanisms, B-Myb and E2F2 can also directly transactivate each other, thus constituting consolidated reciprocal feed-forward transactivation loops. Moreover, both B-Myb and E2F2 are required for the activation of ERK and AKT signaling pathways in colorectal cancer cells. Taken together, our data clarified a critical role for B-Myb in colorectal cancer and unraveled an exquisite mutual collaboration and reciprocal cross regulation between B-Myb and E2F2 that contribute to the malignant progression of human colorectal cancer.

DOI: https://doi.org/10.1038/s41388-021-01961-9
NAR Cancer. 2021 Jun;3(2): zcab019

A novel class of inhibitors that target SRSF10 and promote p53-mediated cytotoxicity on human colorectal cancer cells.

Muhammad Sohail, Lulzim Shkreta, Johanne Toutant, Safwat Rabea, Jean-Philippe Babeu, Caroline Huard, Jasmin Coulombe-Huntington, Aurélie Delannoy, Morgane Placet, Sameh Geha, Fernand-Pierre Gendron, François Boudreau, Mike Tyers, David S Grierson, Benoit Chabot.

The elevated expression of the splicing regulator SRSF10 in metastatic colorectal cancer (CRC) stimulates the production of the pro-tumorigenic BCLAF1-L splice variant. We discovered a group of small molecules with an aminothiazole carboxamide core (GPS167, GPS192 and others) that decrease production of BCLAF1-L. While additional alternative splicing events regulated by SRSF10 are affected by GPS167/192 in HCT116 cells (e.g. in MDM4, WTAP, SLK1 and CLK1), other events are shifted in a SRSF10-independent manner (e.g. in MDM2, NAB2 and TRA2A). GPS167/192 increased the interaction of SRSF10 with the CLK1 and CLK4 kinases, leading us to show that GPS167/192 can inhibit CLK kinases preferentially impacting the activity of SRSF10. Notably, GPS167 impairs the growth of CRC cell lines and organoids, inhibits anchorage-independent colony formation, cell migration, and promotes cytoxicity in a manner that requires SRSF10 and p53. In contrast, GPS167 only minimally affects normal colonocytes and normal colorectal organoids. Thus, GPS167 reprograms the tumorigenic activity of SRSF10 in CRC cells to elicit p53-dependent apoptosis.

DOI: https://doi.org/10.1093/narcan/zcab019
Cell Rep. 2021 Jul 27. pii: S2211-1247(21)00848-2. [Epub ahead of print]36(4): 109431

Identification of cis-HOX-HOXC10 axis as a therapeutic target for colorectal tumor-initiating cells without APC mutations.

Zhenzhen Chen, Jiayi Wu, Benyu Liu, Guangtan Zhang, Zhiwei Wang, Lulu Zhang, Kaili Wang, Zusen Fan, Pingping Zhu.

  Colorectal cancer (CRC) is one of the most common cancers worldwide, in which adenomatous polyposis coli (APC) mutations are frequently and uniquely observed. Here we find that cis-HOX (circular RNA stabilizing HOXC10) is robustly expressed in colorectal tumor-initiating cells (TICs). cis-HOX knockout decreases colorectal TIC numbers and impairs the self-renewal, tumorigenesis, and metastatic capacities of TICs, whereas cis-HOX overexpression drives colorectal TIC self-renewal and metastasis. Mechanistically, cis-HOX binds to HOXC10 mRNA to attenuate its decay through blocking the K-homology splicing regulatory protein (KSRP)-binding sequence of HOXC10 3' UTR. HOXC10 is highly expressed in colorectal tumors and TICs and triggers Wnt/β-catenin activation by activating FZD3 expression. HOXC10 inhibitor salinomycin exerts efficient therapeutic effects in APC-wild-type colorectal tumors, but not in tumors with APC nonsense mutations. Therefore, the cis-HOX-HOXC10 pathway drives colorectal tumorigenesis, stemness, and metastasis and serves as a potential therapeutic target for APC-wild-type colorectal tumors.

Keywords:  APC mutation; FZD3; HOXC10; KSRP; cis-HOX; colorectal cancer; metastasis; self-renewal; tumor initiating cells

DOI:  https://doi.org/10.1016/j.celrep.2021.109431
Cell Biol Int. 2021 Jul 27.

Tazarotene-induced gene 1 interacts with Polo-like kinase 2 and inhibits cell proliferation in HCT116 colorectal cancer cells.

Chun-Hua Wang, Tzung-Ju Lu, Lu-Kai Wang, Chang-Chieh Wu, Mao-Liang Chen, Chan-Yen Kuo, Rong-Yaun Shyu, Fu-Ming Tsai.

  Tazarotene-induced gene 1 (TIG1) is considered to be a tumor suppressor gene that is highly expressed in normal or well-differentiated colon tissues, while downregulation of TIG1 expression occurs in poorly differentiated colorectal cancer (CRC) tissues. However, it is still unclear how TIG1 regulates the tumorigenesis of CRC. Polo-like kinases (Plks) are believed to play an important role in regulating the cell cycle. The performance of PLK2 in CRC is negatively correlated with the differentiation status of CRC tissues. Here, we found that PLK2 can induce the growth of CRC cells and that TIG1 can prevent PLK2 from promoting the proliferation of CRC cells. We also found that the expression of PLK2 in CRC cells was associated with low levels of Fbxw7 protein and increased expression of cyclin E1. When TIG1 was coexpressed with PLK2, the changes in Fbxw7/cyclin E1 levels induced by PLK2 were reversed. In contrast, silencing TIG1 promoted the proliferation of CRC, and when PLK2 was also silenced, the proliferation of CRC cells induced by TIG1 silencing was significantly inhibited. The above research results suggest that TIG1 can regulate the tumorigenesis of CRC by regulating the activity of PLK2. This article is protected by copyright. All rights reserved.

Keywords:  Cell proliferation; Cyclin E1; Fbxw7; Polo-like kinases; Tazarotene-induced gene 1

DOI:  https://doi.org/10.1002/cbin.11681
Cold Spring Harb Perspect Biol. 2021 Jul 26. pii: a040915. [Epub ahead of print]

Insect Gut Regeneration.

Peng Zhang, Bruce A Edgar.

In adult insects, as in vertebrates, the gut epithelium is a highly regenerative tissue that can renew itself rapidly in response to changing inputs from nutrition, the gut microbiota, ingested toxins, and signals from other organs. Because of its cellular and genetic similarities to the mammalian intestine, and its relevance as a target for the control of insect pests and disease vectors, many researchers have used insect intestines to address fundamental questions about stem cell functions during tissue maintenance and regeneration. In Drosophila, where most of the experimental work has been performed, not only are intestinal cell types and behaviors well characterized, but numerous cell signaling interactions have been detailed that mediate gut epithelial regeneration. A prevailing model for regenerative responses in the insect gut invokes stress sensing by damaged enterocytes (ECs) as a principal source for signaling that activates the division of intestinal stem cells (ISCs) and the growth and differentiation of their progeny. However, extant data also reveal alternative mechanisms for regeneration that involve ISC-intrinsic functions, active culling of healthy epithelial cells, enhanced EC growth, and even cytoplasmic shedding by infected ECs. This article reviews current knowledge of the molecular mechanisms involved in gut regeneration in several insect models (Drosophila and Aedes of the order Diptera, and several Lepidoptera).

DOI: https://doi.org/10.1101/cshperspect.a040915
World J Gastroenterol. 2021 Jul 07. 27(25): 3888-3900

Diverse expression patterns of mucin 2 in colorectal cancer indicates its mechanism related to the intestinal mucosal barrier.

Guo-Lian Gan, Hua-Tao Wu, Wen-Jia Chen, Chun-Lan Li, Qian-Qian Ye, Yi-Feng Zheng, Jing Liu.

   BACKGROUND: Abnormal expression patterns of mucin 2 (MUC2) have been reported in a variety of malignant tumors and precancerous lesions. Reduced MUC2 expression in the intestinal mucosa, caused by various pathogenic factors, is related to mechanical dysfunction of the intestinal mucosa barrier and increased intestinal mucosal permeability. However, the relationship between MUC2 and the intestinal mucosal barrier in patients with colorectal cancer (CRC) is not clear.
AIM: To explore the relationship between MUC2 and intestinal mucosal barrier by characterizing the multiple expression patterns of MUC2 in CRC.
METHODS: Immunohistochemical staining was performed on intestinal tissue specimens from 100 CRC patients, including both cancer tissues and adjacent normal tissues. Enzyme-linked immunosorbent assays were performed on preoperative sera from 66 CRC patients and 20 normal sera to detect the serum levels of MUC2, diamine oxide (DAO), and D-lactate (D-LAC). The relationship between MUC2 expression and clinical parameters was calculated by the χ 2 test or Fisher's exact test. Prognostic value of MUC2 was evaluated by Kaplan-Meier curve and log-rank tests.
RESULTS: Immunohistochemical staining of 100 CRC tissues showed that the expression of MUC2 in cancer tissues was lower than that in normal tissues (54% vs 79%, P < 0.05), and it was correlated with tumor-node-metastasis (TNM) stage and lymph node metastasis in CRC patients (P < 0.05). However, the serum level of MUC2 in CRC patients was higher than that in normal controls, and was positively associated with serum levels of human DAO (χ 2 = 3.957, P < 0.05) and D-LAC (χ 2 = 7.236, P < 0.05), which are the biomarkers of the functional status of the intestinal mucosal barrier. And the serum level of MUC2 was correlated with TNM stage, tumor type, and distant metastasis in CRC patients (P < 0.05). Kaplan-Meier curves showed that decreased MUC2 expression in CRC tissues predicted a poor survival.
CONCLUSION: MUC2 in tissues may play a protective role by participating in the intestinal mucosal barrier and can be used as an indicator to evaluate the prognosis of CRC patients.

Keywords:  Colorectal cancer; Expression; Intestinal mucosal barrier; Mucin; Mucin 2; Prognosis

DOI:  https://doi.org/10.3748/wjg.v27.i25.3888
Cancer Sci. 2021 Jul 26.

IGF2BP2 promotes the progression of colorectal cancer through a YAP-dependent mechanism.

Jie Cui, Jiale Tian, Weiwei Wang, Tao He, Xin Li, Chenzheng Gu, Lixin Wang, Jian Wu, Anquan Shang.

   BACKGROUND/AIMS: Insulin-like growth factor 2 (IGF2) mRNA-binding protein 2 (IGF2BP2) is a post-transcriptional regulatory factor that has been implicated in the development of cancer. Herein, this study was performed to examine the possible role of of IGF2BP2 in colorectal cancer (CRC) and its underlying mechanism.
METHODS: First, the expression patterns of IGF2BP2, YAP, and ErbB2 were detected in clinical tissue samples from CRC patients and CRC cell lines. In addition, cell proliferation, migration, invasion and apoptosis were measured and recorded. Enrichment of TEAD4 in the ErbB2 promoter region was measured. Furthermore, IGF2BP2 binding to m6A-modified YAP mRNA was determined using Me-RIP assay with anti-IGF2BP2 antibody. Lastly, a xenograft tumor mouse model was established to substantiate in vitro findings.
RESULTS: IGF2BP2, YAP, and ErbB2 were highly-expressed in CRC tissues and cell lines. In addition, IGF2BP2 was found to be bind to m6A-modified YAP mRNA and increased its stability in CRC cells, thus augmenting the proliferation, migration and invasion of CRC cells while reducing their apoptosis. Meanwhile, YAP regulated ErbB2 expression by promoting TEAD4 enrichment in the ErbB2 promoter region. IGF2BP2 up-regulated ErbB2 by stabilizing YAP expression, and thus promoted proliferation, migration and invasion of CRC cells while attenuating their apoptosis. Also, IGF2BP2 promoted tumor formation in nude mice by regulating the YAP/ErbB2 axis.
CONCLUSION: Overall, our findings indicated that IGF2BP2 may bind to m6A-modified YAP mRNA to promote YAP stability and activate the expression of ErbB2, thus facilitating the progression of CRC.

Keywords:  Apoptosis; Colorectal cancer; ErbB2; IGF2BP2; Invasion; Migration; Proliferation; YAP; m6A

DOI:  https://doi.org/10.1111/cas.15083
Gastroenterol Rep (Oxf). 2021 Jun;9(3): 257-268

lncRNA-PACER upregulates COX-2 and PGE2 through the NF-κB pathway to promote the proliferation and invasion of colorectal-cancer cells.

Peng Sun, Ji-Chuan Quan, Song Wang, Meng Zhuang, Zheng Liu, Xu Guan, Gui-Yu Wang, Hong-Ying Wang, Xi-Shan Wang.

   Background: p50-associated cyclooxygenase-2 extragenic RNA (PACER) is a recently identified antisense long non-coding RNA (lncRNA) located on the upstream of the promoter region of cyclooxygenase-2 (COX-2). Preliminary studies have suggested that PACER is involved in the regulation of COX-2 expression in macrophagocyte and osteosarcoma cells. However, the role of this lncRNA in colorectal cancer (CRC) remains elusive. Here, we investigated the expression of PACER and its effect on cell proliferation and invasion to explore the role of PACER in CRC.
Methods: Real-time quantitative PCR (RT-qPCR) analysis was used to evaluate the expression of PACER in CRC tissues and cells. Methyl thiazolyl tetrazolium (MTT) analysis was then used to investigate the inhibition effect of PACER knock-down in cell proliferation. The promoting role of this lncRNA on invasion by CRC cells was analysed by wound-healing assays, colony-formation assay, and transwell assays. We then used fluorescence in situ hybridization (FISH) to establish the subcellular localization of PACER. COX-2 protein levels were quantified by Western blot analysis and grayscale scanning analysis following the knock-down of PACER. Luciferase assay was carried out to monitor the modulation of the COX-2 promoter region by PACER. Tumor xenografts models were used to investigate the impact of PACER on the tumorigenesis of CRC cells in vivo. Enzyme-linked immunosorbent assay (ELISA) was then used to quantify prostaglandin E2 (PGE2) production upon knock-down of PACER.
Results: RT-qPCR analysis revealed that PACER was highly expressed in CRC tissues and cells, and a high PACER-expression level was associated with poor prognosis. MTT assay, wound-healing assay, colony-formation assay, and transwell assay revealed that PACER enhanced CRC-cell proliferation, invasion, and metastasis in vitro. Analysis of lncRNA localization by FISH showed that it mainly resided in the nucleus. RT-qPCR showed that PACER increased mRNA levels of COX-2. Western blot analysis demonstrated, under normal circumstances, that knock-down of PACER decreased the COX-2 protein level. In the case of p50 absence, COX-2 protein increased rapidly and remained highly expressed after knocking down PACER. Luciferase assay revealed that PACER modulated the COX-2 promoter region. Mouse xenograft models of CRC revealed that PACER promoted colorectal tumorigenesis in vivo. ELISA revealed that PACER knock-down inhibited PGE2 production.
Conclusions: PACER modulates COX-2 expression through the nuclear factor kappa B (NF-κB) pathway in CRC. An increased level of PACER enhances proliferation, migration, and invasion of tumor cells by increasing COX-2 and PGE2 synthesis.

Keywords:  colorectal cancer (CRC); cyclooxygenase-2 (COX-2); lncRNA; p50-associated cyclooxygenase-2 extragenic RNA (PACER)

DOI:  https://doi.org/10.1093/gastro/goaa060
Carcinogenesis. 2021 Jul 27. pii: bgab069. [Epub ahead of print]

Long noncoding RNA MALAT1 sponging miR-26a-5p to modulate Smad1 contributes to colorectal cancer progression by regulating autophagy.

Jiamin Zhou, Miao Wang, Anrong Mao, Yiming Zhao, Longrong Wang, Ye Xu, Hao Jia, Lu Wang.

  Accumulating evidences have suggested that bone morphogenetic protein (BMP) -Smad have a functional role in regulating autophagy in the development of human colorectal cancer (CRC). However, the regulatory mechanisms controlling this process remain unclear. Here, we showed that Smad1, the key effector of BMP2-Smad signaling, induces autophagy by upregulating autophagy-related gene 5 (ATG5) expression, and Smad1 binds to the proximal promoter to induce its expression. Moreover, BMP2 induces autophagy in CRC. Overexpression of Smad1 promotes tumorigenesis and migration of CRC cells, and knockdown of ATG5 is able to rescue the Smad1-induced promotion of CRC proliferation and migration partially. Mechanistically, metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) may act as a competing endogenous RNA by binding with miR-26a-5p competitively and thus modulating the de-repression of downstream target Smad1. Furthermore, clinical analysis results show that Smad1 is positively correlated with MALAT1 and negatively correlated with miR-26a-5p in CRC samples. In conclusion, our results demonstrated that Smad1 may serve as an oncogene in CRC through autophagy.

Keywords:  Autophagy; Colorectal Cancer; MALAT1; Smad1; miR-26a-5p

DOI:  https://doi.org/10.1093/carcin/bgab069
Cancer Genet. 2021 Jul 11. pii: S2210-7762(21)00198-8. [Epub ahead of print]258-259 27-36

Longitudinal change of genetic variations in cetuximab-treated metastatic colorectal cancer.

Sun Young Kim, Kwoneel Kim, Su Han Cho, Sung-Min Chun, Eunyoung Tak, Yong Sang Hong, Jeong Eun Kim, Tae Won Kim.

  Recurrent gene mutations and copy number alterations in cancer patients are presumably associated with resistance to targeted therapy. In the present study, we assessed the gene mutations and copy number alterations that recurrently occurred in cetuximab-treated patients with metastatic colorectal cancer (mCRC). Targeted next-generation sequencing was performed in the tumor samples obtained pre- and postcetuximab treatment to assess the variations that occurred during cetuximab treatment. Moreover, we identified the emergent gene mutations (CDK6, EPHA3, ERCC2, MYC, PCMTD1, PIK3CA, PRIM2, RICTOR, and ZNRF3) and copy number alterations (ARAF, BCL2, BRCA2, EGFR, MYC, and SMAD4) that were recurrently observed only in postprogression samples and not in pretreatment or posttreatment samples from patients revealing clinical response. Furthermore, to identify the feasible candidate variations implicated in treatment resistance, we examined the variants with clonal expansion during treatment and discovered PCBP1 as a variant associated with posttreatment progression. Various recurrent mutations were enriched in the TGF-beta signaling pathway. Collectively, we identified recurrent variations in mCRC samples exhibiting post-cetuximab progression. Additionally, future studies are required to evaluate the therapeutic potential of these variations.

Keywords:  Cetuximab; Colorectal cancer; PCBP1; TGF-beta

DOI:  https://doi.org/10.1016/j.cancergen.2021.06.007
Eur J Pharmacol. 2021 Jul 26. pii: S0014-2999(21)00523-9. [Epub ahead of print] 174370

Diosgenin exerts anti-tumor effects through inactivation of cAMP/PKA/CREB signaling pathway in colorectal cancer.

Si-Yang Li, Jin Shang, Xiao-Mei Mao, Rui Fan, Hui-Qi Li, Rui-Han Li, Dong-Yan Shen.

  Colorectal cancer (CRC) is the most fatal gastrointestinal tumor and it is urge to explore powerful drugs for the treatment. Diosgenin (DSG) as a new steroidal had been reported exerts anti-tumor activity in multiple cancers, including CRC. However, the potential mechanism of DSG suppresses CRC remains further to be revealed. Here, we reported that DSG inhibited proliferation of CRC cells in dose- and time-dependent manner, induced apoptosis by modulating p53 and Bcl-2 family proteins expression to mediate mitochondrial apoptosis pathway, suppressed migration and invasion by reducing MMP-9 (matrix metalloproteinase) and decreased aerobic glycolysis by mediating glucose transporter (GLUT) like GLUT3 and GLUT4, and pyruvate carboxylase PC downregulation. Intriguingly, mechanistic study suggests those phenotypes involved DSG inhibited cAMP/PKA/CREB pathway in CRC cells, and result to inhibit the phosphorylation of CREB to regulate the transcription of genes above-mentioned. Finally, nude mice xenograft tumor model further indicated that DSG could be a great agent to suppress the growth of CRC cells in vivo and have no obvious side effects. Taken together, we revealed a unique mechanism that DSG suppresses CRC cells through cAMP/PKA/CREB pathway and DSG is a promising candidate drug for CRC treatment.

Keywords:  Apoptosis; CREB; Colorectal cancer; Diosgenin; Glycolysis; Motility

DOI:  https://doi.org/10.1016/j.ejphar.2021.174370
Clin Exp Metastasis. 2021 Aug;38(4): 401-410

CXCL2-CXCR2 axis mediates αV integrin-dependent peritoneal metastasis of colon cancer cells.

Mattias Lepsenyi, Nader Algethami, Amr A Al-Haidari, Anwar Algaber, Ingvar Syk, Milladur Rahman, Henrik Thorlacius.

  Peritoneal metastasis is an insidious aspect of colorectal cancer. The aim of the present study was to define mechanisms regulating colon cancer cell adhesion and spread to peritoneal wounds after abdominal surgery. Mice was laparotomized and injected intraperitoneally with CT-26 colon carcinoma cells and metastatic noduli in the peritoneal cavity was quantified after treatment with a CXCR2 antagonist or integrin-αV-antibody. CT-26 cells expressed cell surface chemokine receptors CXCR2, CXCR3, CXCR4 and CXCR5. Stimulation with the CXCR2 ligand, CXCL2, dose-dependently increased proliferation and migration of CT-26 cells in vitro. The CXCR2 antagonist, SB225002, dose-dependently decreased CXCL2-induced proliferation and migration of colon cancer cells in vitro. Intraperitoneal administration of CT-26 colon cancer cells resulted in wide-spread growth of metastatic nodules at the peritoneal surface of laparotomized animals. Laparotomy increased gene expression of CXCL2 at the incisional line. Pretreatment with CXCR2 antagonist reduced metastatic nodules by 70%. Moreover, stimulation with CXCL2 increased CT-26 cell adhesion to extracellular matrix (ECM) proteins in a CXCR2-dependent manner. CT-26 cells expressed the αV, β1 and β3 integrin subunits and immunoneutralization of αV abolished CXCL2-triggered adhesion of CT-26 to vitronectin, fibronectin and fibrinogen. Finally, inhibition of the αV integrin significantly attenuated the number of carcinomatosis nodules by 69% in laparotomized mice. These results were validated by use of the human colon cancer cell line HT-29 in vitro. Our data show that colon cancer cell adhesion and growth on peritoneal wound sites is mediated by a CXCL2-CXCR2 signaling axis and αV integrin-dependent adhesion to ECM proteins.

Keywords:  Chemokines; Chemotaxis; Integrins; Metastasis; Peritoneal carcinomatosis

DOI:  https://doi.org/10.1007/s10585-021-10103-0
Neoplasia. 2021 Jul 25. pii: S1476-5586(21)00056-7. [Epub ahead of print]23(9): 898-911

Trop-2 induces ADAM10-mediated cleavage of E-cadherin and drives EMT-less metastasis in colon cancer.

Emanuela Guerra, Marco Trerotola, Valeria Relli, Rossano Lattanzio, Romina Tripaldi, Giovanna Vacca, Martina Ceci, Khouloud Boujnah, Valeria Garbo, Antonino Moschella, Romina Zappacosta, Pasquale Simeone, Robert de Lange, Ulrich H Weidle, Maria Teresa Rotelli, Arcangelo Picciariello, Raffaella Depalo, Patrizia Querzoli, Massimo Pedriali, Enzo Bianchini, Domenico Angelucci, Giuseppe Pizzicannella, Carla Di Loreto, Mauro Piantelli, Laura Antolini, Xiao-Feng Sun, Donato F Altomare, Saverio Alberti.

  We recently reported that activation of Trop-2 through its cleavage at R87-T88 by ADAM10 underlies Trop-2-driven progression of colon cancer. However, the mechanism of action and pathological impact of Trop-2 in metastatic diffusion remain unexplored. Through searches for molecular determinants of cancer metastasis, we identified TROP2 as unique in its up-regulation across independent colon cancer metastasis models. Overexpression of wild-type Trop-2 in KM12SM human colon cancer cells increased liver metastasis rates in vivo in immunosuppressed mice. Metastatic growth was further enhanced by a tail-less, activated ΔcytoTrop-2 mutant, indicating the Trop-2 tail as a pivotal inhibitory signaling element. In primary tumors and metastases, transcriptome analysis showed no down-regulation of CDH1 by transcription factors for epithelial-to-mesenchymal transition, thus suggesting that the pro-metastatic activity of Trop-2 is through alternative mechanisms. Trop-2 can tightly interact with ADAM10. Here, Trop-2 bound E-cadherin and stimulated ADAM10-mediated proteolytic cleavage of E-cadherin intracellular domain. This induced detachment of E-cadherin from β-actin, and loss of cell-cell adhesion, acquisition of invasive capability, and membrane-driven activation of β-catenin signaling, which were further enhanced by the ΔcytoTrop-2 mutant. This Trop-2/E-cadherin/β-catenin program led to anti-apoptotic signaling, increased cell migration, and enhanced cancer-cell survival. In patients with colon cancer, activation of this Trop-2-centered program led to significantly reduced relapse-free and overall survival, indicating a major impact on progression to metastatic disease. Recently, the anti-Trop-2 mAb Sacituzumab govitecan-hziy was shown to be active against metastatic breast cancer. Our findings define the key relevance of Trop-2 as a target in metastatic colon cancer.

Keywords:  E-cadherin; Epithelial-mesenchymal transition; Metastasis; Signaling networks; Trop-2; proteolytic cleavage; β-catenin

DOI:  https://doi.org/10.1016/j.neo.2021.07.002
JCO Oncol Pract. 2021 Jul 29. OP2100467

Shedding Light on BRAF: Management of Colorectal Cancer in the Era of Personalized Medicine.

Maria Diab.

DOI: https://doi.org/10.1200/OP.21.00467
World J Gastrointest Oncol. 2021 Jul 15. 13(7): 638-661

Systems-level biomarkers identification and drug repositioning in colorectal cancer.

Hande Beklen, Esra Yildirim, Medi Kori, Beste Turanli, Kazim Yalcin Arga.

  Colorectal cancer (CRC) is the most commonly diagnosed fatal cancer in both women and men worldwide. CRC ranked second in mortality and third in incidence in 2020. It is difficult to diagnose CRC at an early stage as there are no clinical symptoms. Despite advances in molecular biology, only a limited number of biomarkers have been translated into routine clinical practice to predict risk, prognosis and response to treatment. In the last decades, systems biology approaches at the omics level have gained importance. Over the years, several biomarkers for CRC have been discovered in terms of disease diagnosis and prognosis. On the other hand, a few drugs are being developed and used in clinics for the treatment of CRC. However, the development of new drugs is very costly and time-consuming as the research and development takes about 10 years and more than $1 billion. Therefore, drug repositioning (DR) could save time and money by establishing new indications for existing drugs. In this review, we aim to provide an overview of biomarkers for the diagnosis and prognosis of CRC from the systems biology perspective and insights into DR approaches for the prevention or treatment of CRC.

Keywords:  Biomarker; Colon cancer; Colorectal cancer; Drug repositioning; Omics; Systems biology

DOI:  https://doi.org/10.4251/wjgo.v13.i7.638
Cancer Cell Int. 2021 Jul 28. 21(1): 402

Expression and prognostic significance of CBX2 in colorectal cancer: database mining for CBX family members in malignancies and vitro analyses.

He Zhou, Yong Xiong, Zuoliang Liu, Songlin Hou, Tong Zhou, Yongfu Xiong.

   BACKGROUND: The Chromobox (CBX) domain protein family, a core component of polycomb repressive complexes 1, is involved in transcriptional repression, cell differentiation, and program development by binding to methylated histone tails. Each CBX family member plays a distinct role in various biological processes through their own specific chromatin domains, due to differences in conserved sequences of the CBX proteins. It has been demonstrated that colorectal cancer (CRC) is a multiple-step biological evolutionary process, whereas the roles of the CBX family in CRC remain largely unclear.
METHODS: In the present study, the expression and prognostic significance of the CBX family in CRC were systematically analyzed through a series of online databases, including Cancer Cell Line Encyclopedia (CCLE), Oncomine, Human Protein Atlas (HPA), and Gene Expression Profiling Interactive Analysis (GEPIA). For in vitro verification, we performed cell cloning, flow cytometry and transwell experiments to verify the proliferation and invasion ability of CRC cells after knocking down CBX2.
RESULTS: Most CBX proteins were found to be highly expressed in CRC, but only the elevated expression of CBX2 could be associated with poor prognosis in patients with CRC. Further examination of the role of CBX2 in CRC was performed through several in vitro experiments. CBX2 was overexpressed in CRC cell lines via the CCLE database and the results were verified by RT-qPCR. Moreover, the knockdown of CBX2 significantly suppressed CRC cell proliferation and invasion. Furthermore, the downregulation of CBX2 was found to promote CRC cell apoptosis.
CONCLUSIONS: Based on these findings, CBX2 may function as an oncogene and potential prognostic biomarker. Thus, the association between the abnormal expression of CBX2 and the initiation of CRC deserves further exploration.

Keywords:  CBX2; Cell proliferation; Chromobox; Colorectal cancer; Prognosis

DOI:  https://doi.org/10.1186/s12935-021-02106-4
Cancer Drug Resist. 2021 ;4 684-709

Cancer stem cells, epithelial-mesenchymal transition, ATP and their roles in drug resistance in cancer.

Haiyun Zhang, Alexander Steed, Milo Co, Xiaozhuo Chen.

  The cancer stem cell (CSC) state and epithelial-mesenchymal transition (EMT) activation are tightly interconnected. Cancer cells that acquire the EMT/CSC phenotype are equipped with adaptive metabolic changes to maintain low reactive oxygen species levels and stemness, enhanced drug transporters, anti-apoptotic machinery and DNA repair system. Factors present in the tumor microenvironment such as hypoxia and the communication with non-cancer stromal cells also promote cancer cells to enter the EMT/CSC state and display related resistance. ATP, particularly the high levels of intratumoral extracellular ATP functioning through both signaling pathways and ATP internalization, induces and regulates EMT and CSC. The three of them work together to enhance drug resistance. New findings in each of these factors will help us explore deeper into mechanisms of drug resistance and suggest new resistance-associated markers and therapeutic targets.

Keywords:  ABC transporters; ATP internalization; Tumor microenvironment; apoptosis; biological markers; macropinocytosis

DOI:  https://doi.org/10.20517/cdr.2021.32
Int J Cancer. 2021 Jul 26.

Mutational profiles of metastatic colorectal cancer treated with FOLFIRI plus cetuximab or bevacizumab before and after secondary resection (AIO KRK 0306; FIRE-3).

Arndt Stahler, Volker Heinemann, Julian Walter Holch, Jobst Christian von Einem, Christoph Benedikt Westphalen, Kathrin Heinrich, Laura Schlieker, Ivan Jelas, Annabel Helga Sophie Alig, Laura Elisabeth Fischer, Lena Weiss, Dominik Paul Modest, Ludwig Fischer von Weikersthal, Thomas Decker, Alexander Kiani, Markus Moehler, Florian Kaiser, Thomas Kirchner, Andreas Jung, Sebastian Stintzing.

  Secondary resection of metastases is recommended in metastatic colorectal cancer (mCRC). Data describing changes in mutational profiles of corresponding primary tumor and metastatic tissue after conversion treatment are limited. Next generation sequencing was performed in formalin-fixed mCRC samples from patients of the FIRE-3 trial (FOLFIRI plus cetuximab or bevacizumab) before treatment start (baseline) and after secondary resection of metastases (post baseline). Changes of mutational profiles and tumor mutational burden (TMB) were assessed within a post-hoc analysis. Median overall survival (OS), progression-free survival (PFS) and objective response rate (ORR) were compared between treatment arms. Paired tumor samples were obtained from 25 patients (19 RAS wild-type, 6 RAS mutant by pyrosequencing). ORR (92.0% vs 58.0%) and OS (60.8 vs 35.4 months, hazard ratio = 0.39 [95% CI 0.14-1.12], P = .08) were higher for patients receiving cetuximab. After conversion therapy, 56 alterations (42 in the cetuximab and 14 in the bevacizumab arm) were newly observed in 18 patients (9 each treated with cetuximab or bevacizumab). Gains (n = 21) and losses (n = 21) of alterations occurred during cetuximab-based treatment, while mainly gains of alterations occurred during bevacizumab (n = 10). Three of nine patients treated with cetuximab that presented a change of mutational profiles, developed resistance to cetuximab. Mutational profiles were largely comparable before and after treatment with anti-VEGF or anti-EGFR directed monoclonal antibodies after secondary resection. Mutations associated with resistance to anti-EGFR antibodies were observed in only one-third of patients.

Keywords:  NGS; bevacizumab; cetuximab; metastatic colorectal cancer; paired samples

DOI:  https://doi.org/10.1002/ijc.33747
Cancer Sci. 2021 Jul 30.

Impact of UGT1A1 Genotype on the Efficacy and Safety of Irinotecan-based Chemotherapy in Metastatic Colorectal Cancer.

Satoru Iwasa, Kei Muro, Satoshi Morita, Young Suk Park, Masato Nakamura, Masahito Kotaka, Tomohiro Nishina, Hiroshi Matsuoka, Joong Bae Ahn, Keun-Wook Lee, Yong Sang Hong, Sae Won Han, Sang-Hee Cho, Dong-Sheng Zhang, Wei-Jia Fang, Li Bai, Xiang-Lin Yuan, Ying Yuan, Yasuhide Yamada, Junichi Sakamoto, Tae Won Kim.

  The phase III AXEPT study demonstrated the non-inferiority of modified capecitabine plus irinotecan (mXELIRI) with or without bevacizumab relative to fluorouracil, leucovorin, and irinotecan (FOLFIRI) with or without bevacizumab as a second-line treatment for metastatic colorectal cancer (mCRC). We evaluated the associations between the UGT1A1 genotype linked to adverse events - caused by irinotecan - and the efficacy and safety of mXELIRI and FOLFIRI. The UGT1A1 genotype was prospectively determined and patients were categorized into 3 groups according to wild-type (WT, *1/*1), single heterozygous (SH, *28/*1 or *6/*1), and double heterozygous or homozygous (DHH, *28/*28, *6/*6, or *28/*6). Overall survival (OS), progression-free survival, response rate, and safety were assessed. The UGT1A1 genotype was available in all 650 randomized patients (WT: 309 [47.5%]; SH: 291 [44.8%]; DHH: 50 [7.7%]). The median OS was 15.9, 17.7, and 10.6 months in the WT, SH, and DHH groups, respectively, with an adjusted hazard ratio (HR) of 1.53 (95% confidence interval [CI] 1.12-2.09; P = 0.008) for DHH vs. WT or SH. The median OS in the mXELIRI and FOLFIRI arms was 18.1 vs. 14.3 months (HR 0.80; 95% CI 0.62-1.03) in the WT group, 16.3 vs. 18.3 months (HR 1.04; 95% CI 0.79-1.36) in the SH group, and 13.0 vs. 9.1 months (HR 0.71; 95% CI 0.39-1.31) in the DHH group, respectively. mXELIRI with or without bevacizumab could be a standard second-line chemotherapy in terms of efficacy and safety regardless of the UGT1A1 genotype.

Keywords:   UGT1A1 ; XELIRI; capecitabine; colorectal cancer; irinotecan

DOI:  https://doi.org/10.1111/cas.15092
J Crohns Colitis. 2021 Jul 26. pii: jjab134. [Epub ahead of print]

An IRF1-dependent pathway of TNFα-induced shedding in intestinal epithelial cells.

Gao Tan, Chongyang Huang, Jiaye Chen, Bingxia Chen, Yanqiang Shi, Fachao Zhi.

   BACKGROUND: Shedding of intestinal epithelial cells (IECs) is a potent cause of barrier loss that plays an important role in the pathogenesis of inflammatory bowel disease (IBD). TNFα can induce IEC shedding, but little is known about this process.
METHODS: To investigate the molecular mechanism regulating IEC shedding, mice lacking interferon regulatory factor1 (IRF1), caspase-3 or gasdermin E (GSDME) and their control wild-type (WT) littermates were intravenously injected with TNFα to establish an IEC shedding model. A dual-luciferase reporter assay and a chromatin immunoprecipitation assay were used to determine the role of IRF1 in regulating caspase-3 expression.
RESULTS: TNFα administration induced obvious IEC shedding in WT mice, but IRF1 -/- and caspase-3 -/-mice were completely protected from TNFα-induced IEC shedding. As a critical transcription factor, IRF1 was found to be required for caspase-3 expression in IECs by binding to IRF1-binding sites in the caspase-3 promoter. In WT mice, plasma membrane integrity was disrupted in shed IECs; these cells were swollen and contained GSDME-N terminal (NT) fragments, which are responsible for the induction of pyroptosis. However, in Gsdme -/- mice, plasma membrane integrity was not disrupted in shed IECs, which were not swollen and did not contain GSDME-NT, indicating that GSDME converted TNFα-induced IEC shedding into a pyroptotic cell death process. In addition, IRF1 deficiency resulted in decreases in mucosal inflammation and mucosal bacteria levels in TNFα-challenged colons.
CONCLUSIONS: IRF1 deficiency maintains intestinal barrier integrity by restricting TNFα-induced IEC shedding.

Keywords:  Epithelial shedding; Inflammatory bowel disease; Interferon regulatory factor 1; Intestinal epithelial cell; Pyroptosis

DOI:  https://doi.org/10.1093/ecco-jcc/jjab134
DNA Repair (Amst). 2021 Jul 09. pii: S1568-7864(21)00134-8. [Epub ahead of print]106 103178

Tissue-specific reduction in MLH1 expression induces microsatellite instability in intestine of Mlh1+/- mice.

Kul S Shrestha, Elli-Mari Aska, Minna M Tuominen, Liisa Kauppi.

  Tumors of Lynch syndrome (LS) patients display high levels of microsatellite instability (MSI), which results from complete loss of DNA mismatch repair (MMR), in line with Knudson's two-hit hypothesis. Why some organs, in particular those of the gastrointestinal (GI) tract, are prone to tumorigenesis in LS remains unknown. We hypothesized that MMR is haploinsufficient in certain tissues, compromising microsatellite stability in a tissue-specific manner before tumorigenesis. Using mouse genetics, we tested how levels of MLH1, a central MMR protein, affect age- and tissue-specific microsatellite stability in vivo and whether elevated MSI is detectable prior to loss of MMR function and to neoplastic growth. To assess putative tissue-specific MMR haploinsufficiency, we determined relevant molecular phenotypes (MSI, Mlh1 promoter methylation status, MLH1 protein and RNA levels) in jejuna of Mlh1+/- mice and compared them to those in spleen, as well as to MMR-proficient and -deficient controls (Mlh1+/+ and Mlh1-/- mice). While spleen MLH1 levels of Mlh1+/- mice were, as expected, approximately 50 % compared to wildtype mice, MLH1 levels in jejunum varied substantially between individual Mlh1+/- mice and moreover, decreased with age. Mlh1+/- mice with soma-wide Mlh1 promoter methylation often displayed severe MLH1 depletion in jejunum. Reduced (but still detectable) MLH1 levels correlated with elevated MSI in Mlh1+/- jejunum. MSI in jejunum increased with age, while in spleens of the same mice, MLH1 levels and microsatellites remained stable. Thus, MLH1 expression levels are particularly labile in intestine of Mlh1+/- mice, giving rise to tissue-specific MSI long before neoplasia. A similar mechanism likely also operates also in the human GI epithelium and could explain the wide range in age-of-onset of LS-associated tumorigenesis.

Keywords:  Lynch syndrome; Microsatellite instability; Mlh1 haploinsufficiency; Mlh1 promoter methylation

DOI:  https://doi.org/10.1016/j.dnarep.2021.103178
Future Oncol. 2021 Jul 29.

The prognostic and predictive impact of BRAF mutations in deficient mismatch repair/microsatellite instability-high colorectal cancer: systematic review/meta-analysis.

Robin Park, Laercio Lopes, Sunggon Lee, Ivy Riano, Anwaar Saeed.

  Aims: The authors present a systematic review/meta-analysis of the impact of BRAF mutations on prognosis and immune checkpoint inhibitor (ICI) response in deficient mismatch repair/microsatellite instability-high colorectal cancer. Methods: Hazard ratios for overall survival and odds ratios for objective response rate to ICIs were calculated in BRAF-mutated versus BRAF wild-type patients. Results: After screening, nine and three studies, respectively, were included for analysis of prognosis (analysis A) and ICI response (analysis B). Analysis A showed worse overall survival in BRAF-mutated compared with BRAF wild-type stage I-IV patients (hazard ratio: 1.57; 95% CI: 1.23-1.99), and analysis B showed no difference in objective response rate (odds ratio: 1.04; 95% CI: 0.48-2.25). Conclusion: BRAF mutations are associated with worse overall survival but not differential response to ICIs in deficient mismatch repair/microsatellite instability-high colorectal cancer.

Keywords:  BRAF; immunotherapy; microsatellite instability; mismatch repair deficiency; prognosis

DOI:  https://doi.org/10.2217/fon-2021-0552
Bioorg Chem. 2021 Jul 16. pii: S0045-2068(21)00548-4. [Epub ahead of print]115 105171

Discovery of small molecule acting as multitarget inhibitor of colorectal cancer by simultaneous blocking of the key COX-2, 5-LOX and PIM-1 kinase enzymes.

Mostafa M M El-Miligy, Ahmed K Al-Kubeisi, Saad R El-Zemity, Rasha A Nassra, Marwa M Abu-Serie, Aly A Hazzaa.

  Colorectal cancer (CRC) is the second cause of cancer death worldwide. Inhibitors of COX-2, 5-LOX and PIM-1 kinase were very effective in the treatment and prevention of CRC in mouse models in vivo. Furthermore, thymol was confirmed to inhibit CRC cell proliferation in cancer cell lines and inhibitory activity against COX-2 and 5-LOX. On the other hand, 4-thiazolidinone pharmacophore was incorporated in the structures of various reported COX-2, 5-LOX and PIM kinase inhibitors. Consequently, the aim of the present investigation was to combat CRC by synthesis and biological evaluation of new thymol - 4-thiazolidinone hybrids as multitarget anticancer agents that could inhibit the key COX-2, 5-LOX and PIM-1 kinase enzymes simultaneously. Compounds 5a-d and 5g displayed inhibitory activity against COX-2 nearly equal to Celecoxib with high selectivity index (SI). Moreover, compounds 5b-e showed 5-LOX inhibitory activity nearly equal to the reference Quercetin while compounds 5a, 5f and 5g elicited inhibitory activity slightly lower than Quercetin. Furthermore, in vivo formalin-induced paw edema test revealed that, compounds 5a, 5c, 5f and 5g showed higher % inhibition than Celecoxib and compounds 5a, 5f and 5g showed higher % inhibition than Diclofenac sodium. In addition, compounds 5a-c, 5e-g showed in vivo superior gastrointestinal safety profile as Celecoxib in fasted rats. Besides, compounds 5d, 5e and 5g exhibited the highest activity against human CRC cell lines (Caco-2 and HCT-116) at doses less than their EC100 on normal human cells. Furthermore, compounds 5e and 5g induced apoptosis-dependent death by above 50% in the treated CRC cell lines. Moreover, compounds 5e and 5g induced caspase activation by >50% in human CRC. Also, compounds 5d, 5e and 5g showed in vitro inhibitory activity against both PIM-1\2 kinases comparable to the reference Staurosporine. In silico docking studies were concordant with the biological results. In conclusion, compound 5g, of simple chemical structure, achieved the target goal of inhibiting three targets leading to inhibition of human CRC cell proliferation. It inhibited the target key enzymes COX-2, 5-LOX and PIM-1\2 kinase in vitro. Besides, it revealed in vitro inhibition of cell proliferation in cancer cell lines via activation of caspase 3\7 dependent-apoptosis in human CRC cell lines. In addition, it elicited in vivo anti-inflammatory activity in formalin-induced paw edema test and in vivo oral safety in gastric ulcerogenic activity test.

Keywords:  COX; CRC; LOX; PIM kinase; Thiazolidinone; Thymol

DOI:  https://doi.org/10.1016/j.bioorg.2021.105171
Front Pharmacol. 2021 ;12 673432

Oxymatrine Synergistically Enhances Doxorubicin Anticancer Effects in Colorectal Cancer.

Di Pan, Wen Zhang, Nenling Zhang, Yini Xu, Yi Chen, Jianqing Peng, Yan Chen, Yanyan Zhang, Xiangchun Shen.

  The combination of chemotherapy with natural products is a common strategy to enhance anticancer effects while alleviating the dose-dependent adverse effects of cancer treatment. Oxymatrine (OMT) has been extensively reported as having anticancer activity. Doxorubicin (DOX) is a chemotherapeutic DNA-damaging agent used for the treatment of carcinoma. In this study, we investigated whether synergistic effects exist with the combination treatment with OMT and DOX using human colorectal cancer cell (CRC) lines and the potential mechanisms involved in in vitro and in vivo activities. The MTT and colony formation assay results showed that compared to either OMT or DOX monotherapy, the combination of OMT + DOX markedly inhibited the growth of HT-29 and SW620 cells. Wound healing assays showed significant inhibition of cell migration with co-treatment, supported by the change in E-cadherin and N-cadherin expressions in Western blotting. Furthermore, flow cytometry analysis revealed that OMT + DOX co-treatment enhanced cell apoptosis as a result of ROS generation, whereas NAC attenuated OMT + DOX-induced apoptosis. Similarly, the apoptosis-related proteins (cleaved caspase-3, cleaved caspase-9, and the ratio of Bax/Bcl-2) were determined by Western blotting, which showed that the expressions of these markers were notably increased in the co-treatment group. Furthermore, co-administration of a low dose of DOX and OMT inhibited xenograft tumor growth in a dose-dependent manner. TUNEL assay and Ki67 staining images indicated more apoptosis and less proliferation occurred in OMT plus DOX-treated xenograft tumors. Meanwhile, the combination strategy decreased cardiotoxicity, which is the most serious side effect of DOX. RNA sequencing was performed to explore the precise molecular alterations involved in the combination group. Among the numerous differentially expressed genes, downregulated FHL-2 and upregulated cleaved SPTAN1 were validated in both mRNA and protein levels of HT-29 and SW620 cells. These two proteins might play a pivotal role involving in OMT + DOX synergistic activity. Overall, OMT in combination with DOX presented an outstanding synergistic antitumor effect, indicating that this beneficial combination may offer a potential therapy for CRC patients.

Keywords:  RNA-seq; colorectal cancer; doxorubicin; oxymatrine; synergistically effect

DOI:  https://doi.org/10.3389/fphar.2021.673432
Trends Cell Biol. 2021 Jul 22. pii: S0962-8924(21)00140-9. [Epub ahead of print]

Cell cycle on the crossroad of tumorigenesis and cancer therapy.

Jing Liu, Yunhua Peng, Wenyi Wei.

  Aberrancy in cell cycle progression is one of the fundamental mechanisms underlying tumorigenesis, making regulators of the cell cycle machinery rational anticancer therapeutic targets. A growing body of evidence indicates that the cell cycle regulatory pathway integrates into other hallmarks of cancer, including metabolism remodeling and immune escape. Thus, therapies against cell cycle machinery components can not only repress the division of cancer cells, but also reverse cancer metabolism and restore cancer immune surveillance. Besides the ongoing effects on the development of small molecule inhibitors (SMIs) of the cell cycle machinery, proteolysis targeting chimeras (PROTACs) have recently been used to target these oncogenic proteins related to cell cycle progression. Here, we discuss the rationale of cell cycle targeting therapies, particularly PROTACs, to more efficiently retard tumorigenesis.

Keywords:  PROTAC; cancer; cancer immune; cell cycle; degradation; metabolism

DOI:  https://doi.org/10.1016/j.tcb.2021.07.001