染色质重构复合体蛋白PBRM1对膀胱癌细胞生物学功能的影响

Effects of chromatin-remodeling complex PBRM1 on biological features of bladder cancer cells

目的探讨染色质重构复合体蛋白PBRM1对膀胱癌细胞株EJ、T24细胞生物学功能的影响。方法针对PBRM1 mRNA序列设计合成siRNA,转染膀胱癌细胞,沉默PBRM1基因后进行如下检测:RT-PCR法检测PBRM1 mRNA表达变化及凋亡、迁移相关指标;MTT法检测细胞增殖;流式细胞术检测细胞凋亡情况;Western印迹法检测凋亡蛋白Caspase3表达水平;划痕实验及Matrigel实验检测细胞迁移及侵袭能力变化。结果 PBRM1siRNA能有效抑制膀胱癌细胞中PBRM1的表达。PBRM1基因沉默后,与空白对照组相比,细胞增殖能力显著增强,同时细胞凋亡减少,细胞迁移及侵袭能力明显减弱。凋亡及迁移相关基因mRNA水平及蛋白水平也出现相应变化。结论 PBRM1对膀胱癌细胞株EJ、T24生物学功能有重要影响,可抑制细胞增殖,促进细胞凋亡、迁移及侵袭。

Objective To investigate the effect of silencing PBRM1 gene by siRNA on biological features of bladder cancer cells. Methods PBRMl-specific siRNA was designed and synthesized. The siRNA was transfected into bladder cancer cells EJ and T24. After silencing PBRM1 gene, the mRNA expressions of apoptosis and migration-related genes were detected by RT-PCR; cell growth inhibition was determined by MTT; cell apoptosis was assayed by flow cytometry （FCM）; expression of caspase3 protein was detected by Western blot; migration and invasion was detected by scratch test and Matrigel test. Results PBRMl-specific siRNA efficiently suppressed the PBRM1 expression in bladder cancer cells. After bladder cancer cells were transfected with PBRMI-siRNA, cell proliferation rate was increased; the percentage of apoptotic cells was decreased; cell migration and invasion was inhibited. The mRNA expression of apoptosis-related genes Caspase8, Caspase3, Bid, Bcl-2 and migration-related genes E-cadherin, MMP9, MMP2, Vime and their protein levels were also changedaccordingly. Conclusion The biological functions of PBRM1 in bladder cancer cell line EJ and T24 are involved in inhibiting cell growth and enhancing cell apoptosis, metastasis and invasion.