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Circ_0005962 functions as an oncogene to aggravate non-small cell lung cancer progression via circ_0005962/miR-382-5p/PDK4 regulatory network

By Zhihong Zhang, Zhenxiu Shan, Rubin Chen, Xiaorong Peng, Bin Xu, Liang Xiao, Guofei Zhang

Posted 06 Mar 2020
bioRxiv DOI: 10.1101/2020.03.06.980185

Non-small cell lung cancer (NSCLC) is a leading threat to human lives with high incidence and mortality. Circular RNAs (circRNAs) were reported to play important roles in human cancers. The purpose of this study was to investigate the role of circ\_0005962 and explore the underlying functional mechanisms. The expression of circ\_0005962, miR-382-5p and pyruvate dehydrogenase kinase 4 (PDK4) was detected by quantitative real-time polymerase chain reaction (qRT-PCR). Cell proliferation and cell apoptosis were assessed by cell counting kit-8 (CCK-8) assay and flow cytometry assay, respectively. The protein levels of Beclin 1, light chain3 (LC3-II/LC3-I), PDK4, Cleaved Caspase 3 (C-caspase 3) and proliferating cell nuclear antigen (PCNA) were examined using western blot analysis. Glycolysis was determined according to the levels of glucose consumption and lactate production. The interaction between miR-382-5p and circ\_0005962 or PDK4 was predicted by the online tool CircInteractome or starbase and verified by dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay. Xenograft model was constructed to investigate the role of circ\_0005962 in vivo. circ\_0005962 expressed with a high level in NSCLC tissues and cells. Circ\_0005962 knockdown inhibited proliferation, autophagy, and glycolysis but promoted apoptosis in NSCLC cells. MiR-382-5p was targeted by circ\_0005962, and its inhibition reversed the role of circ\_0005962 knockdown. Besides, PDK4, a target of miR-382-5p, was regulated by circ\_0005962 through miR-382-5p, and its overexpression abolished the effects of miR-382-5p reintroduction. Circ\_0005962 knockdown suppressed tumor growth in vivo. Circ\_0005962 knockdown restrained cell proliferation, autophagy, and glycolysis but stimulated apoptosis through modulating the circ\_0005962/miR-382-5p/PDK4 axis. Our study broadened the insights into understanding the mechanism of NSCLC progression.

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