沉默KLF11表达通过Wnt/β-catenin信号通路对胃癌细胞BGC823增殖和凋亡的影响

Effect of specific small interfering RNA against KLF11 expression on proliferation and apoptosis of human gastric cancer cell line BGC823 via Wnt/β-catenin signaling pathway

目的探讨KLF11在胃癌组织中的表达及其对胃癌细胞BGC823增殖和凋亡的影响。方法收集60例胃癌组织及其对应的癌旁组织,用RT-PCR检测KLF11 mRNA表达,通过小分子RNA干扰抑制胃癌细胞BGC823中KLF11表达水平;MTT法检测细胞的活力;流式细胞仪检测细胞周期、凋亡率;采用Western blot检测细胞周期、凋亡相关指标蛋白和Wnt/β-catenin的改变;采用酶标仪检测Caspase3酶活性。结果KLF11 mRNA在胃癌组织相对表达水平较癌旁组织升高(t=11.38,P<0.05),其表达与肿瘤大小、浸润深度、淋巴结转移及TNM分期均有关(均P<0.05);沉默KLF11表达后,胃癌BGC823细胞的增殖能力受抑制(F=19.56,P<0.05);G 0/G 1期细胞数目增多[(41.40%±0.98%)比(66.53%±1.01%),F=32.69,P<0.05],并且凋亡率升高[(41.44%±1.59%)比(15.42%±0.86%),F=35.35,P<0.05];Bax、Cleaved Caspase3蛋白表达上升(F=23.33、33.63,均P<0.05),β-catenin、Bcl-2、CyclinD1、CyclinE蛋白表达降低(F=22.21、16.24、26.75、33.42,均P<0.05),Caspase3酶的活性增加(F=16.56,P<0.05)。结论KLF11在胃癌组织中呈高表达,下调其表达可抑制Wnt/β-catenin信号通路,抑制胃癌细胞增殖,诱导细胞凋亡。

Objective To investigate the expression of KLF11 in gastric cancer tissues and cell lines as well as its effect on proliferation and apoptosis of human gastric cancer cells BGC823.Methods Sixty pairs gastric cancer tissues and corresponding adjacent tissues were collected.The expression of KLF11 mRNA in gastric cancer tissues and their adjacent tissues was detected by RT-PCR.The expression of KLF11 was detected in gastric cancer cells.KLF11 expression was silenced.The proliferation of cells were detected by using MTT assay.Flow cytometry was used to detect the cell cycle and cell apoptosis rate.Western bloting was used to examine the changes of concentration of proteins associated with cell cycle,cell apoptosis and Wnt/β-catenin signaling pathway related proteins.The activity of Caspase3 enzyme was detected by spectrophotometry.Results The relative expression of KLF11 mRNA in gastric carcinoma tissues was significantly higher than that of the adjacent tissues(t=11.38,P<0.05).Its expression was related to tumor size,depth of invasion,lymph node metastasis and TNM clinical stage(all P<0.05).The proliferation of BGC823 cells was significantly suppressed after KLF11 silencing(F=19.56,P<0.05),and the cell cycle was arrested in G0/G1 phase[(41.40%±0.98%)vs.(66.53%±1.01%),F=32.69,P<0.05].Meanwhile,the apoptosis rate was significantly increased by KFL11 silencing[(41.44%±1.59%)vs.(15.42%±0.86%),F=35.35,P<0.05].The results of Western blotting revealed that the expression of Bax and Cleaved Caspase3 was significantly increased(F=23.33,33.63;both P<0.05),wheras that ofβ-catenin,Bcl-2,CyclinD1and CyclinE was significantly reduced(F=22.21,16.24,26.75,33.42;all P<0.05).The activity of Caspase3 enzyme was enhanced(F=16.56,P<0.05).Conclusion KLF11 was highly-expressed in gastric cancer tissues and cells,KFL11 silencing could inhibit gastric cancer cells proliferation and induce cell apoptosis via Wnt/β-catenin signaling pathway.