bims-meluca 2020-01-05 papers

on Metabolism of non-small cell lung carcinoma

Issue of 2020–01–05
six papers selected by
the Muñoz-Pinedo/Nadal (PReTT) lab, L’Institut d’Investigació Biomèdica de Bellvitge and Cristina Muñoz Pinedo, L’Institut d’Investigació Biomèdica de Bellvitge

Erastin/sorafenib induces cisplatin-resistant non-small cell lung cancer cell ferroptosis through inhibition of the Nrf2/xCT pathway.
Distinct Roles of VEGFA and ANGPT2 in Lung Adenocarcinoma and Squamous Cell Carcinoma.
Genome-wide global identification of NRF2 binding sites in A549 non-small cell lung cancer cells by ChIP-Seq reveals NRF2 regulation of genes involved in focal adhesion pathways.
Metabolic and lipidomic characterization of malignant pleural effusion in human lung cancer.
Independent prognostic value of HIF-1α expression in radiofrequency ablation of lung cancer.
PRDX5 as a novel binding partner in Nrf2-mediated NSCLC progression under oxidative stress.

Oncol Lett. 2020 Jan;19(1): 323-333

Erastin/sorafenib induces cisplatin-resistant non-small cell lung cancer cell ferroptosis through inhibition of the Nrf2/xCT pathway.

Yu Li, Hengyi Yan, Xiaoman Xu, Hongbo Liu, Cen Wu, Li Zhao.

  Non-small cell lung cancer (NSCLC) has long been one of the most lethal types of cancer due to its lack of typical clinical symptoms at early stages and high risk of tumour recurrence, even following complete surgical resection. Multicourse chemotherapy based on cisplatin (CDDP) is the standard adjuvant treatment for NSCLC; however, its benefits for the overall survival of patients are limited. In this study, NSCLC cells possessing CDDP-resistant characteristics (N5CP cells), obtained from surgical resection of clinical specimens of patients with NSCLC, were cultured and screened to generate research models. This study aimed to identify the mechanism underlying tumour cell resistance to CDDP and to identify a novel treatment for NSCLC following CDDP failure. CDDP-mediated NF-E2 related factor 2 (Nrf2)/light chain of System xc - (xCT) pathway activation was associated with the resistance of cells to CDDP. Therefore, erastin/sorafenib regulation of Nrf2 or xCT expression may alter the sensitivity of tumour cells to CDDP. The small molecules erastin and sorafenib effectively induced N5CP cell ferroptosis, which was mediated by the accumulation of intracellular lipid reactive oxygen species. Additionally, low doses of erastin or sorafenib could be used in association with CDDP to effectively trigger N5CP cell ferroptosis. Furthermore, it was indicated that erastin and sorafenib, alone or in combination with a low dose of CDDP, effectively inhibited the growth of N5CP cells in vivo. Therefore, ferroptosis inducers, including erastin and sorafenib, may be considered a novel treatment regimen for patients with NSCLC, particularly patients with CDDP failure.

Keywords:  NF-E2 related factor 2 pathway; cisplatin resistance; erastin; ferroptosis; non-small cell lung cancer cell; sorafenib

DOI:  https://doi.org/10.3892/ol.2019.11066
J Cancer. 2020 ;11(1): 153-167

Distinct Roles of VEGFA and ANGPT2 in Lung Adenocarcinoma and Squamous Cell Carcinoma.

Shuang Qin, Ming Yi, Dechao Jiao, Anping Li, Kongming Wu.

  Background: Vascular endothelial growth factor A (VEGFA) and angiopoietin 2 (ANGPT2) are key mediators in angiogenesis. The expression and clinical significance of VEGFA and ANGPT2 have been investigated in lung cancer, but the results are controversial. The specific roles of VEGFA and ANGPT2 in adenocarcinoma (ADC) and squamous cell carcinoma (SQC) are still not fully understood. To characterize it, we conducted the current study. Materials and Methods: The relationships between clinic-pathological characteristics and the protein expressions of VEGFA and ANGPT2 were analyzed on tissue microarrays by immunohistochemistry (IHC) staining. Then public databases were used to evaluate the association of VEGFA and ANGPT2 mRNA expressions with clinic-pathological parameters and prognosis. Cobalt chloride (CoCl2) was adopted to mimic a hypoxic microenvironment and western blot was used to detect the expression of hypoxia inducible factor-1α (HIF-1α), VEGFA and ANGPT2 in lung cancer cell lines. Results: IHC staining revealed that the expressions of VEGFA and ANGPT2 were enriched in lung cancer tissues compared with normal tissues. Additionally, both VEGFA and ANGPT2 protein levels were significantly associated with the tumor size and lymph node metastasis only in ADC, not SQC. More importantly, increased VEGFA and ANGPT2 protein levels were negatively correlated with overall survival (OS) of ADC individuals. Meta-analyses of 22 gene expression omnibus (GEO) databases of lung cancer implicated that patients with higher VEGFA and ANGPT2 mRNA expressions tended to have advanced stage in ADC rather than SQC. Kaplan-Meier plot analyses further verified that high levels of VEGFA and ANGPT2 mRNA were associated with poor survival only in ADC. Moreover, the combination of VEGFA and ANGPT2 could more precisely predict prognosis in ADC. In hypoxia-mimicking conditions, induced expression of HIF-1α unregulated VEGFA and ANGPT2 proteins abundance. Conclusion: Our results showed hypoxia upregulated the protein levels of VEGFA and ANGPT2 in lung cancer cell lines, and the roles of VEGFA and ANGPT2 were distinct in ADC and SQC. Combined detections of VEGFA and ANGPT2 may be valuable prognostic biomarkers for ADC and double block of VEGFA and ANGPT2 may improve therapeutic outcome.

Keywords:  ADC; ANGPT2; NSCLC; SQC; VEGFA; prognosis

DOI:  https://doi.org/10.7150/jca.34693
Aging (Albany NY). 2019 Dec 28. 11(24): 12600-12623

Genome-wide global identification of NRF2 binding sites in A549 non-small cell lung cancer cells by ChIP-Seq reveals NRF2 regulation of genes involved in focal adhesion pathways.

Akhileshwar Namani, Kaihua Liu, Shengcun Wang, Xihang Zhou, Yijiao Liao, Hongyan Wang, Xiu Jun Wang, Xiuwen Tang.

  Nuclear factor erythroid-derived-2-like 2(NRF2) regulates its downstream genes through binding with antioxidant responsive elements in their promoter regions. Hyperactivation of NRF2 results in oncogenesis and drug resistance in various cancers including non-small cell lung cancer (NSCLC). However, identification of the genes and pathways regulated by NRF2 in NSCLC warrants further investigation. We investigated the global NRF2 genomic binding sites using the high-throughput ChIP-Seq technique in KEAP1 (Kelch-like ECH-associated protein 1)-mutated A549 (NSCLC) cells. We next carried out an integrated analysis of the ChIP-Seq data with transcriptomic data from A549 cells with NRF2-knockdown and RNA-Seq data from TCGA patients with altered KEAP1 to identify downstream and clinically-correlated genes respectively. Furthermore, we applied transcription factor enrichment analysis, generated a protein-protein interaction network, and used kinase enrichment analysis. Moreover, functional annotation of NRF2 binding sites using DAVID v7 identified the genes involved in focal adhesion. Putative focal adhesion genes regulated by NRF2 were validated using qRT-PCR. Further, we selected one novel conserved focal adhesion gene regulated by NRF2-LAMC1 (laminin subunit gamma 1) and validated it using a reporter assay. Overall, the identification of NRF2 target genes paves the way for identifying the molecular mechanism of NRF2 signaling in NSCLC development and therapy. Moreover, our data highlight the complexity of the pathways regulated by NRF2 in lung tumorigenesis.

Keywords:  ChIP-seq; focal adhesion; microarray; non-small-cell lung cancer; nuclear factor erythroid-derived-2-like 2

DOI:  https://doi.org/10.18632/aging.102590
J Pharm Biomed Anal. 2019 Dec 23. pii: S0731-7085(19)31859-X. [Epub ahead of print]180 113069

Metabolic and lipidomic characterization of malignant pleural effusion in human lung cancer.

Zhiyi Yang, Zhengbo Song, Zhongjian Chen, Zhenyu Guo, Hangbiao Jin, Cheng Ding, Yanjun Hong, Zongwei Cai.

  Malignant pleural effusion (MPE) is an important hallmark for late-stage lung cancer with metastasis. Current clinical diagnosis methods require tedious work to distinguish MPE from benign pleural effusion (BPE). The objective of this study was to characterize the metabolic signatures in MPE of lung cancer, and identify potential metabolite biomarkers for diagnosis of MPE. MPE from lung cancer (n = 46) and BPE from tuberculosis patients (n = 32) were investigated by liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based global metabolomic and lipidomic profiling. Multivariate partial least-square discriminative analysis models exhibited distinct metabolic profiles between MPE and BPE. A total of 25 ether lipids, including phosphatidylcholines (PC), lysophosphatidylcholines (LPC) and phosphatidylethanolamines (PE), were observed to be significantly downregulated in MPE with excellent diagnostic potential. Plasmalogen PC(40:3p) showed highest AUC value of 0.953 in receiver operating characteristic (ROC) model. Oxidized polyunsaturated fatty acids (PUFA) were upregulated in MPE. The obtained results implied a high oxidative stress and peroxisome disorder in lung cancer patients. Combined metabolomic and lipidomic profiling have discovered potential biomarkers in MPE with excellent clinical diagnostic capability. Dysregulated ether lipids and oxidized PUFAs have implied an aberrant redox metabolism, which provides novel insights into the pathology of MPE in lung cancer.

Keywords:  Biomarker; Lipidomics; Lung cancer; Mass spectrometry; Metabolomics; Pleural effusion

DOI:  https://doi.org/10.1016/j.jpba.2019.113069
Oncol Lett. 2020 Jan;19(1): 849-857

Independent prognostic value of HIF-1α expression in radiofrequency ablation of lung cancer.

Jun Wan, Xiean Ling, Zhanpeng Rao, Bin Peng, Guanggui Ding.

  Radiofrequency ablation (RFA) is widely used in the treatment of lung cancer. Hypoxia-inducible factor-1α (HIF-1α) is a crucial transcription factor regulating oxygen homeostasis that is involved in tumor cell metastasis. The present study investigated the impact of HIF-1α expression and other factors, such as postoperative blood CD4+/CD8+ ratio, on the prognosis of patients with lung cancer who had received RFA treatment. A total of 80 patients with lung cancer were recruited between January 2011 and October 2016 at The Shenzhen People's Hospital. Lung cancer was confirmed following pathological or histological examination. All patients underwent RFA treatment. Patients were followed up for 6-66 months. HIF-1α expression in lung cancer tissues was assessed by immunohistochemistry. Multivariate survival analysis was performed using Cox proportional hazards model. The results demonstrated that HIF-1α level was low in 36 patients and overexpressed in 44 patients with lung cancer. Kaplan-Meier (KM) curve analysis demonstrated that the overall survival time of patients with high HIF-1α expression was significantly shorter compared with patients with low HIF-1α expression (P<0.05). Furthermore, the results from the KM model and log-rank test revealed that age, Union for International Cancer Control stage, primary or metastatic cancer, chemotherapy, postoperative blood CD4+/CD8+ ratio, Eastern Cooperative Oncology Group performance status and HIF-1α expression had significant effects on overall survival of patients with lung cancer. The results from Cox analysis demonstrated that high HIF-1α expression, advanced age, clinical staging and chemotherapy were independent risk factors for the prognosis of lung cancer following RFA treatment, and that high HIF-1α expression was associated with the increased risk (5.91-fold) of mortality. In conclusion, the present study demonstrated that HIF-1α expression was increased in lung cancer tissues and was associated with the prognosis of patients with lung cancer who were treated with RFA. These findings suggest that HIF-1α expression may be considered as a marker for evaluating the prognosis of these patients.

Keywords:  hypoxia-inducible factor-1α; lung cancer; prognosis; radiofrequency ablation

DOI:  https://doi.org/10.3892/ol.2019.11130
Aging (Albany NY). 2020 Jan 03. 11

PRDX5 as a novel binding partner in Nrf2-mediated NSCLC progression under oxidative stress.

Xinming Chen, Xiang Cao, Weizhang Xiao, Ben Li, Qun Xue.

  Non-small-cell lung cancer (NSCLC) is one of the most common malignant tumors in the world. Reactive oxidative species (ROS) and nuclear factor-related factor 2 (Nrf2) -antioxidant response element (ARE) signal pathway are known to play important roles in the development of NSCLC. In this study, we identified Peroxiredoxin 5 (PRDX5) as a novel binding partner for Nrf2. PRDX5 was significantly increased in human NSCLC specimens and cell lines. Nrf2 interacted with PRDX5 in H2O2-stimulated NCSLC cells, and the interaction promoted the expression of NAD(P)H: quinone oxidoreductase 1 (NQO1) protein in NSCLC cells. Further, high expression of Nrf2 and PRDX5 were associated with worsened prognosis in patients with NSCLC significantly. Moreover, animal studies showed that the growth of tumors treated with Nrf2 and PRDX5 shRNA was significantly lower than that of the other groups. All these data indicated that overexpressed PRDX5 in NSCLC promoted binding with Nrf2 and enhanced NQO1 expression and NSCLC development. Overall, our studies demonstrated that PRDX5 can be a novel binding partner of Nrf2 in promoting NCSLC development under oxidative stress and provide potential opportunity for improving NSCLC therapy.

Keywords:  NSCLC; nuclear factor-related factor 2; oxidative stress; peroxiredoxin 5

DOI:  https://doi.org/10.18632/aging.102605