Rab5 GTPase对膀胱癌细胞自噬及增殖的调控

目的探讨Rab5 GTPase对膀胱癌细胞自噬及增殖的影响。方法采用实时定量PCR检测60例膀胱癌组织以及相应正常膀胱组织中Rab5 mRNA的表达水平,并检测Rab5 mRNA在膀胱癌细胞系及正常膀胱细胞系中的表达情况。通过pcDNA3.1-Rab5转染膀胱癌细胞系BIU-87、RT4过表达Rab5;免疫荧光、Western blot检测自噬相关蛋白LC3Ⅱ/Ⅰ、LC3斑点数目变化情况以及自噬相关蛋白Vps34的表达变化;采用CCK-8检测膀胱癌细胞的增殖改变情况。结果Rab5 mRNA在膀胱癌组织中的表达高于正常膀胱组织,Rab5 mRNA在膀胱癌细胞中的表达高于正常膀胱细胞系,差异有统计学意义(P<0.05);免疫荧光显示过表达Rab5后LC3荧光斑点明显增多,且过表达Rab5促进膀胱癌细胞中Vps34的表达增加及LC3-Ⅰ向LC3-Ⅱ的转化;过表达Rab5能促进膀胱癌细胞增殖(P<0.05)。结论膀胱癌组织中Rab5 mRNA的表达高于正常膀胱组织,Rab5提高膀胱癌细胞自噬水平并促进膀胱癌细胞增殖,且与自噬相关蛋白Vps34相关。

The regulation of Rab5 GTPase on autophagy and cells proliferation in bladder cancer cells

Objective To explore the regulation of Rab5 GTPase on autophagy and cell proliferation in bladder cancer cell.Methods Real-time PCR was used to measure the expression of Rab5 mRNA in bladder cancer tissues and adjacent normal tissues.The expression of Rab5 was overexpressed in BIU-87 and RT4 cell lines by pcDNA3.1-Rab5.Immunofluorescence was used to analyze the spots number of LC3,Western blot was used to detect the expression of LC3Ⅱ/Ⅰand Vps34.The proliferation level of bladder cancer cells was analyzed by CCK-8.Results The expression level of Rab5 mRNA was up-regulated in bladder cancer tissues than in the adjacent normal tissues.The expression of Rab5 mRNA in bladder cancer cells was higher than that in normal bladder cells.Overexpressed Rab5 could promote autophagy and cells proliferation in bladder cancer cells.The expression of Vps34 and LC3 II could be enhanced by overexpressed Rab5.Conclusion Rab5 could promote autophagy and proliferation in bladder cancer cells.The promotion of Rab5 on autophagy might be related to Vps34.