MIR-145 REGULATES THE BIOLOGICAL BEHAVIOR OF NEPHROBLASTOMA SKNEP-1 CELLS BY INHIBITING THE PI3K / AKT SIGNALING PATHWAY

Liang Gan^#, Wenbo Wu^#, Ning Ding, Jing Tang, Pinde Rao, Zhifeng Duan, Qiang Tao^*

Department of Urology, The Affiliated Children’s Hospital of Nanchang University, Jiangxi Provincial Children’s Hospital, Nanchang 330006, Jiangxi Province, China

Objective: To analyze the regulatory effect of microRNA-145 (miR-145) on the biological behavior of nephroblastoma sknep-1 cells by inhibiting the phosphatidylinositol 3 kinase/protein kinase B (PI3K/Akt) signal pathway.

Methods: Tumoral tissue from 28 cases of Wilms’ tumor was collected from April 2019 to February 2020; para-tumoral tissue from the same individuals was chosen as a control. Additionally, miR-145 was overexpressed in cultured Sknep-1 cells using lentivirus infection; an empty viral vector was used for the cell culture controls. The tissue expression of miR-145 was measured in all samples using real-time quantitative PCR. Cell proliferation was determined using cell counting Kit-8, and apoptosis was detected using acridine orange/ethidium bromide fluorescence staining. Transwell cell migration assay was used to determine the migratory response of the cultured cells. Expression levels of Bax, Bcl-2, Caspase-3, PI3K, and Akt were detected using Western blot.

Results: The expression of miR-145 was significantly lower in the tumor tissue than in control tissue samples (P<.01). Expression levels of miR-145, Bcl-2, caspase-3, and apoptosis rate in miR-145 were significantly increased in cells overexpressing miR-145 compared to controls. Conversely, Bax, PI3K, and Akt expression levels, together with cell proliferation and migration ability, were significantly decreased in the overexpressing group (P<.01).

Conclusion: Overexpression of miR-145 may inhibit the proliferation and migration of Wilms’ tumor and promote cell apoptosis by inhibiting the activation of the PI3K/Akt signaling pathway.