沉默S100P对膀胱癌5637细胞生物学特性的影响
Effect of S100P gene silencing on biological properties of bladder cancer 5637 cell

目的:探讨S100P基因对膀胱癌细胞生物学特性的影响。方法:将膀胱癌5637细胞分为空白对照组、阴性对照组和S100P siRNA组。空白对照组常规培养,S100P siRNA组细胞感染含S100P siRNA的慢病毒,阴性对照组感染空病毒。感染48 h后,采用实时荧光定量PCR检测3组细胞中S100P mRNA的表达水平,采用Western blot法检测S100P、Cyclin D1、CDK2、P53、NF-κB、Ezrin、P-gp蛋白的表达水平,MTT法检测细胞增殖抑制率及吡柔比星对细胞的IC50,流式细胞术检测细胞凋亡率。结果:与空白对照组和阴性对照组比较,S100P siRNA组细胞S100P mRNA和蛋白表达水平,Cyclin D1、CDK2、P53、NF-kB、Ezrin、P-gp蛋白表达水平均降低(P<0.05)。S100P siRNA组细胞增殖抑制率和凋亡率均高于阴性对照组(P<0.05),吡柔比星的IC50低于空白对照组(P<0.05)。结论:S100P与膀胱癌细胞的增殖、凋亡及化疗敏感性关系密切。
Aim: To study the effect of S100P gene silencing on the biological properties and chemosensitivity of human bladder cell.Methods: The bladder cell 5637 cells were divided into three groups: the blank control group, the negative control group and S100P siRNA group. The cells in S100P siRNA group were infected with S100P siRNA lentivirus. The negative control group was infected with empty virus. After 48 h, the mRNA and protein of S100P,and Cyclin D1, CDK2, P53, NF-κB, Ezrin and P-gp proteins in cells were detected by qRT-PCR and Western blot,respectively. The proliferation inhibition rate, the half inhibitory concentration(IC50)of pirarubicin and cell apoptosis rates were detected by MTT assay and flow cytometry.Results: The expressions of S100P mRNA and protein in S100P siRNA group were significantly lower than those in the blank control group and negative control group(P<0.05), as well as the protein expressions of Cyclin D1, CDK2, P53, NF-κB, Ezrin and P-gp. The apoptosis rate and proliferation inhibition rate of S100P siRNA group were higher than negative control group, and IC50 of pirarubicin was significantly lower than the blank control group(P<0.05).Conclusion: S100P gene is correlated with the proliferation,apoptosis and chemosensitivity of bladder cancer