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  

Effect of PKM2 on the invasive ability of bladder urothelial carcinoma cells by regulating the reactive oxygen species-dependent PI3K/Akt signaling pathway

作　　者：梁凯[1] 殷磊[2] 陈琦 LIANG Kai;YIN Lei;CHEN Qi(Department of Urology,Heilongjiang Provincal Hospital,Harbin 150001,China;Department of Urology,The First Hospital of China Medical University,Shenyang 110001,China)

机构地区：[1]黑龙江省医院泌尿外科,哈尔滨150001 [2]中国医科大学附属第一医院泌尿外科,沈阳110001

出　　处：《中国医科大学学报》2024年第7期621-627,634,共8页Journal of China Medical University

基　　金：辽宁省科技厅应用基础研究计划[辽科发(2022) 43号]。

摘　　要：目的 探讨PKM2通过调控活性氧(ROS)依赖的PI3K/Akt信号通路对膀胱尿路上皮细胞癌细胞侵袭能力的影响。方法 将人膀胱癌T24细胞随机分为Control组、si-NC组、si-PKM2组和si-PKM2+IGF-1组。实时PCR检测细胞中PKM2 mRNA表达;CCK-8检测细胞增殖;Transwell小室法检测细胞侵袭和转移;Hoechst 33342荧光染色和流式细胞术检测细胞凋亡;ROS荧光探针法检测细胞中ROS水平;Western blotting检测细胞中PI3K、Akt、mTOR、caspase-3、Bax、Bcl-2蛋白表达。结果 与人正常膀胱上皮SV-HUC-1细胞相比,人膀胱癌T24细胞中PKM2 mRNA相对表达量明显增加(P <0.05)。与Control组相比,si-PKM2组T24细胞中PKM2 mRNA相对表达量、细胞增殖能力、侵袭细胞数以及细胞中p-PI3K/PI3K、p-Akt/Akt、p-mTOR/mTOR比值和Bcl-2蛋白表达水平明显降低(P <0.05),细胞凋亡率、细胞中ROS相对水平以及细胞中Bax、caspase-3蛋白表达水平明显升高(P <0.05);与siPKM2组相比,si-PKM2+IGF-1组T24细胞中PKM2 mRNA相对表达量、细胞增殖能力、侵袭细胞数以及细胞中p-PI3K/PI3K、p-Akt/Akt、p-mTOR/mTOR比值和Bcl-2蛋白表达水平明显升高(P <0.05),细胞凋亡率、细胞中ROS相对水平以及细胞中Bax、caspase-3蛋白表达水平明显降低(P <0.05)。结论 敲减PKM2可促进膀胱尿路上皮细胞癌细胞中ROS水平升高,抑制PI3K/Akt/mTOR信号通路激活,进而发挥抑制细胞侵袭和促进细胞凋亡作用。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.PKM2 mRNA 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 cytometry,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 expression of PKM2 mRNA,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 PKM2 mRNA,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 PKM2 can 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.

关 键 词：M2型丙酮酸激酶 膀胱尿路上皮细胞癌 活性氧 PI3K/Akt/mTOR信号通路 

分 类 号：R737.14[医药卫生—肿瘤]