Effect of PKM2 on the invasive ability of bladder urothelial carcinoma cells by regulating the reactive oxygen species-dependent PI3K/Akt signaling pathway
10.12007/j.issn.0258-4646.2024.07.008
- VernacularTitle:PKM2通过调控活性氧依赖的PI3K/Akt信号通路影响膀胱尿路上皮细胞癌细胞的侵袭能力
- Author:
Kai LIANG
1
;
Lei YIN
;
Qi CHEN
Author Information
1. 黑龙江省医院泌尿外科,哈尔滨 150001
- Keywords:
pyruvate kinase M2;
bladder urothelial carcinoma;
reactive oxygen species;
PI3K/Akt/mTOR signaling pathway
- From:
Journal of China Medical University
2024;53(7):621-627,634
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effect of PKM2 on the invasive ability of bladder urothelial carcinoma cells by regulating the reactive oxygen species(ROS)-dependent PI3K/Akt signaling pathway.Methods Human bladder cancer T24 cells were divided into the control,si-NC,si-PKM2,and si-PKM2+IGF-1 groups.PKM2mRNA expression,proliferation,invasion and migration,and apoptosis of T24 cells were detected using real-time PCR,CCK-8 assay,Transwell assay,and Hoechst 33342 fluorescence staining and flow cyto-metry,respectively.The ROS level in the cells was measured using ROS fluorescence probe staining.Western blotting was used to detect the expression of PI3K,Akt,mTOR,caspase-3,Bax,and Bcl-2 proteins.Results The expression of PKM2 mRNA in human bladder cancer T24 cells was significantly higher than in normal bladder SV-HUC-1 cells(P<0.05).Compared to the control group,the expres-sion of PKM2mRNA,cell proliferation ability,number of invading cells,p-PI3K/PI3K ratio,p-Akt/Akt ratio,p-mTOR/mTOR ratio,and Bcl-2 protein expression in T24 cells in the si-PKM2 group were significantly decreased(P<0.05),whereas the apoptosis rate,relative ROS level,and expression of Bax and caspase-3 proteins were significantly increased(P<0.05).Compared to the si-PKM2 group,the expression of PKM2mRNA,cell proliferation ability,number of invading cells,p-PI3K/PI3K ratio,p-Akt/Akt ratio,p-mTOR/mTOR ratio,and Bcl-2 protein expression in T24 cells in the si-PKM2+IGF-1 group were significantly increased(P<0.05),whereas the apoptosis rate,relative ROS level,and expression of Bax and caspase-3 proteins were significantly decreased(P<0.05).Conclusion Knockdown of PKM2can increase the level of ROS in bladder urothelial carcinoma cells and inhibit the activation of the PI3K/Akt/mTOR signaling pathway,thus inhibiting cell invasion and promoting cell apoptosis.
