Role of microRNA in Regulating Cell Signaling Pathways, Cell Cycle, and Apoptosis in Non-Small Cell Lung Cancer

2016 ◽  
Vol 16 (5) ◽  
pp. 474-486 ◽  
Author(s):  
D.K. Singh ◽  
S. Bose ◽  
S. Kumar
2021 ◽  
Author(s):  
Yi Liao ◽  
Jianguo Feng ◽  
Weichao Sun ◽  
Chao Wu ◽  
Jingyao Li ◽  
...  

Abstract Background: Cold-inducible RNA binding protein (CIRP) is a newly discovered proto-oncogene. In this study, we investigated the role of CIRP in the progression of non-small cell lung cancer (NSCLC) using clinic samples, cultured cell lines and animal lung cancer models. Methods: Tissue arrays, IHC and HE staining, immunoblotting, and qRT-PCR were used to detect the indicated gene expression; Plasmid and siRNA transfections as well as viral infection were used to manipulate gene expression; Cell proliferation assay, cell cycle analysis, cell migration and invasion analysis, soft agar colony formation assay, tail intravenous injecting and subcutaneously inoculating of animal models were performed to study the role of CIRP in NSCLC cells; Gene expression microarray was used to select the underlying pathways; RNA immunoprecipitation assay, biotin pull-down assay, immuno-purification assay, mRNA decay analyses and luciferase reporter assay were performed to elucidate the mechanisms. The log-rank (Mantel-Cox) test, independent sample T test, the nonparametric Mann-Whitney test, spearman rank test and two-tailed independent sample T-test were used accordingly in our study. Results: Our data showed that CIRP was highly expressed in NSCLC tissue, and its level was negatively correlated with the prognosis of NSCLC patients. By manipulating CIRP expression in A549, H460, H1299, and H1650 cell lines, we demonstrated that CIRP overexpression promoted the transition of G1/G0 phase to S phase and the formation of enhanced malignant phenotype of NSCLC, reflected by increased proliferation, enhanced invasion/metastasis and greater tumorigenic capabilities both in vitro and in vivo. Transcriptome sequencing further demonstrated that CIRP acted on cell cycle, DNA replication and Wnt signaling pathway to exert its pro-oncogenic action. Mechanistically, CIRP directly bound to the 3’- and 5'-UTR of CTNNB1 mRNA, leading to enhanced stability and translation of CTNNB1 mRNA and promote IRES-mediated protein synthesis, respectively. Eventually, the increased CTNNB1 protein levels mediated excessive activation of the Wnt/β-Catenin signaling pathway and its downstream C-myc, COX-2, CCND1, MMP7, VEGFA and CD44. Conclusion: Our results support CIRP as a candidate oncogene in NSCLC and a potential target for NSCLC therapy.


Biology ◽  
2020 ◽  
Vol 9 (8) ◽  
pp. 201 ◽  
Author(s):  
Shweta Arora ◽  
Prithvi Singh ◽  
Arshad Husain Rahmani ◽  
Saleh A. Almatroodi ◽  
Ravins Dohare ◽  
...  

Lung cancer is a prime cause of worldwide cancer deaths, with non-small cell lung cancer (NSCLC) as a frequent subtype. Surgical resection, chemotherapy are the currently used treatment methods. Delayed detection, poor prognosis, tumor heterogeneity, and chemoresistance make them relatively ineffective. Genomic medicine is a budding aspect of cancer therapeutics, where miRNAs are impressively involved. miRNAs are short ncRNAs that bind to 3′UTR of target mRNA, causing its degradation or translational repression to regulate gene expression. This study aims to identify important miRNA-mRNA-TF interactions in NSCLC using bioinformatics analysis. GEO datasets containing mRNA expression data of NSCLC were used to determine differentially expressed genes (DEGs), and identification of hub genes-BIRC5, CCNB1, KIF11, KIF20A, and KIF4A (all functionally enriched in cell cycle). The FFL network involved, comprised of miR-20b-5p, CCNB1, HMGA2, and E2F7. KM survival analysis determines that these components may be effective prognostic biomarkers and would be a new contemplation in NSCLC therapeutics as they target cell cycle and immunosurveillance mechanisms via HMGA2 and E2F7. They provide survival advantage and evasion of host immune response (via downregulation of cytokines-IL6, IL1R1 and upregulation of chemokines-CXCL13, CXCL14) to NSCLC. The study has provided innovative targets, but further validation is needed to confirm the proposed mechanism.


2020 ◽  
Author(s):  
Yanlong Yang ◽  
Xiaobo Chen ◽  
Hongwen Sun ◽  
Qinghe Yu ◽  
Fang Yin ◽  
...  

Abstract Background: The role of Centromere protein O (CENPO) in the development of non-small cell lung cancer (NSCLC) is unknown. This study aimed to investigate the potential roleof CENPO in the development of NSCLCMethods: The expression level of CENPO was investigated in both tissues and cell-lines. Celigo cell counting assay, wound-healing assay, Transwell assay and Flow cytometry was used to explore the effect of CENPO on proliferation, migration, invasion, apoptosis and cell cycle of NSCLC. The potential mechanism of CENPO was explored by Human Apoptosis Antibody, also, western blot was conducted to detect the expression of PI3K/Akt/mTOR pathway and cell-cycle related protein (mTOR, P-mTOR, CDK1, CDK6 and PIK3CA). Besides, the effect of CENPO on the growth of NSCLC solid tumors was demonstrated in vivo.Results: Our study suggested CENPO gene overexpression in NSCLC. Reduced CENPO expression substantially decreased the proliferation, migration and invasion ability, and promoting apoptosis and induces cell cycle arrest of NSCLC cell-lines. Preliminary mechanism research suggested reduced CENPO could active apoptosis pathway, suppressing PI3K/AKT/mTOR pathway and down-regulation cell-cycle related proteins.Conclusion: CENPO was up-regulated in NSCLC and played an important role in promoting the progress of NSCLC.


Author(s):  
Yi Liao ◽  
Jianguo Feng ◽  
Weichao Sun ◽  
Chao Wu ◽  
Jingyao Li ◽  
...  

Abstract Background Cold-inducible RNA binding protein (CIRP) is a newly discovered proto-oncogene. In this study, we investigated the role of CIRP in the progression of non-small cell lung cancer (NSCLC) using patient tissue samples, cultured cell lines and animal lung cancer models. Methods Tissue arrays, IHC and HE staining, immunoblotting, and qRT-PCR were used to detect the indicated gene expression; plasmid and siRNA transfections as well as viral infection were used to manipulate gene expression; cell proliferation assay, cell cycle analysis, cell migration and invasion analysis, soft agar colony formation assay, tail intravenous injection and subcutaneous inoculation of animal models were performed to study the role of CIRP in NSCLC cells; Gene expression microarray was used to select the underlying pathways; and RNA immunoprecipitation assay, biotin pull-down assay, immunopurification assay, mRNA decay analyses and luciferase reporter assay were performed to elucidate the mechanisms. The log-rank (Mantel-Cox) test, independent sample T-test, nonparametric Mann-Whitney test, Spearman rank test and two-tailed independent sample T-test were used accordingly in our study. Results Our data showed that CIRP was highly expressed in NSCLC tissue, and its level was negatively correlated with the prognosis of NSCLC patients. By manipulating CIRP expression in A549, H460, H1299, and H1650 cell lines, we demonstrated that CIRP overexpression promoted the transition of G1/G0 phase to S phase and the formation of an enhanced malignant phenotype of NSCLC, reflected by increased proliferation, enhanced invasion/metastasis and greater tumorigenic capabilities both in vitro and in vivo. Transcriptome sequencing further demonstrated that CIRP acted on the cell cycle, DNA replication and Wnt signaling pathway to exert its pro-oncogenic action. Mechanistically, CIRP directly bound to the 3′- and 5′-UTRs of CTNNB1 mRNA, leading to enhanced stability and translation of CTNNB1 mRNA and promoting IRES-mediated protein synthesis, respectively. Eventually, the increased CTNNB1 protein levels mediated excessive activation of the Wnt/β-catenin signaling pathway and its downstream targets C-myc, COX-2, CCND1, MMP7, VEGFA and CD44. Conclusion Our results support CIRP as a candidate oncogene in NSCLC and a potential target for NSCLC therapy.


2020 ◽  
Author(s):  
Yanlong Yang ◽  
Xiaobo Chen ◽  
Hongwen Sun ◽  
Qinghe Yu ◽  
Fang Yin ◽  
...  

Abstract Background: The role of Centromere protein O (CENPO) in the development of non-small cell lung cancer (NSCLC) is unknown. This study aimed to investigate the potential roleof CENPO in the development of NSCLCMethods: The expression level of CENPO was investigated in both tissues and cell-lines. Celigo cell counting assay, wound-healing assay, Transwell assay and Flow cytometry was used to explore the effect of CENPO on proliferation, migration, invasion, apoptosis and cell cycle of NSCLC. The potential mechanism of CENPO was explored by Human Apoptosis Antibody, also, western blot was conducted to detect the expression of PI3K/Akt/mTOR pathway and cell-cycle related protein (mTOR, P-mTOR, CDK1, CDK6 and PIK3CA). Besides, the effect of CENPO on the growth of NSCLC solid tumors was demonstrated in vivo.Results: Our study suggested CENPO gene overexpression in NSCLC. Reduced CENPO expression substantially decreased the proliferation, migration and invasion ability, and promoting apoptosis and induces cell cycle arrest of NSCLC cell-lines. Preliminary mechanism research suggested reduced CENPO could active apoptosis pathway, suppressing PI3K/AKT/mTOR pathway and down-regulation cell-cycle related proteins.Conclusion: CENPO was up-regulated in NSCLC and played an important role in promoting the progress of NSCLC.


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