PRR11在膀胱癌组织中的表达及其对膀胱癌T24细胞增殖和凋亡的影响

Expression of PRR11 in Bladder Cancer Tissue and Its Effect on Proliferation and Apoptosis of Bladder Cancer Cell Line T24

目的探讨富含脯氨酸蛋白11(PRR11)在膀胱癌组织中的表达及其基因沉默对膀胱癌T24细胞增殖和凋亡的影响。方法采用免疫组织化学法检测57例膀胱尿路上皮癌组织及其癌旁组织中PRR11蛋白的表达,并分析PRR11蛋白表达水平与患者临床病理特征的关系。qRT-PCR和Western blot检测人永生化膀胱上皮细胞株SV-HUC-1以及人膀胱癌细胞株HTB-9、T24、J82和UM-UC-3细胞中PRR11 mRNA和蛋白表达水平。慢病毒shRNA干扰技术沉默膀胱癌T24细胞中PRR11基因表达,qRT-PCR检测细胞中PRR11 mRNA表达水平;CCK-8检测细胞增殖活性;平板克隆形成实验检测细胞克隆形成能力;流式细胞术检测细胞凋亡率;Western blot检测细胞中PRR11、Caspase-3、Bcl-2和Bax的表达水平。结果PRR11在膀胱尿路上皮癌组织中高表达,并与患者肿瘤病理分级和T分期显著相关。HTB-9、T24、J82和UM-UC-3细胞中PRR11 mRNA和蛋白表达水平较SV-HUC-1细胞显著升高(P<0.05),其中T24细胞最为显著。PRR11基因沉默后,T24细胞中PRR11 mRNA和蛋白表达水平以及细胞增殖活性和克隆形成能力显著降低、细胞凋亡率增高、Cleaved-caspase-3和Bax蛋白表达上调,Bcl-2蛋白表达下调。结论PRR11在膀胱尿路上皮癌组织及膀胱癌细胞株中高表达,干扰PRR11表达可抑制膀胱癌T24细胞增殖,并促进其细胞凋亡。

Objective To investigate the expression of PRR11 in bladder cancer tissues and its effect on proliferation and apoptosis of bladder cancer cell line T24.Methods The expression of PRR11 was detected using immunohistochemistry method in 57 specimens of bladder urothelial carcinoma and adjacent tissues.The correlations of PRR11 expression with the clinicopathological characteristics of patients with bladder urothelial carcinoma were analyzed.The mRNA and protein expression levels of PRR11 in human immortalized bladder epithelial cell lines SV-HUC-1 and human bladder cancer cell lines HTB-9,T24,J82 and UM-UC-3 were measured by qRT-PCR and Western blot.The gene expression of PRR11 in T24 cells was silenced by lentivirus shRNA.The mRNA expression level of PRR11 was detected by qRT-PCR.CCK-8 was used to detect cell proliferative activity.Cell clonality was detected by plate cloning assays.The rate of apoptosis was evaluated using flow cytometry.The protein expression levels of PRR11,Caspase-3,Bcl-2 and Bax were assessed by Western blot.Results PRR11 was highly expressed in bladder urothelial carcinoma,and its expression level was correlated with the pathological grade and T stage of the tumor.The mRNA and protein expression levels of PRR11 in HTB-9,T24,J82 and UM-UC-3 cells were higher than those in SV-HUC-1 cells(P<0.05),especially in T24 cells.PRR11 gene silence reduced the expression levels of PRR11 mRNA and protein,as well as the cell proliferation activity and cell clonality,elevated the apoptosis rate,up-regulated the protein expression levels of Cleaved caspase-3 and Bax,and down-regulated the protein expression level of Bcl-2.Conclusion The expression of PRR11 is upregulated in bladder urothelial carcinoma tissues and bladder cancer cell lines.Interfering with PRR11 expression can inhibit the proliferation and promote the apoptosis of bladder cancer T24 cells.