SIRT5基因沉默对结肠癌细胞的迁移和侵袭能力的影响

Effects of SIRT5 Gene Silencing on Migration and Invasion of Colonic Carcinoma Cells

目的探讨沉默信息调节因子5(silent information regulator 5,SIRT5)基因沉默对结肠癌细胞的迁移和侵袭能力的影响。方法逆转录聚合酶链反应(reverse transcription-polymerase chain reaction,RT-PCR)及Western blot分析SIRT5在多种结肠癌细胞系和永生化结肠上皮细胞中的表达。Western blot检测基质金属蛋白酶2(matrix metalloproteinase,MMP2)及基质金属蛋白酶9(matrix metalloproteinase,MMP 9)、上皮细胞-间充质转化(epithelial mesenchymal transition,EMT)、指标蛋白[上皮型钙黏蛋白(E-cadherin)、神经型钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)]的表达。结果 SIRT5在结肠癌细胞系中较人正常结肠上皮细胞比较,蛋白表达水平明显上调,尤其是结肠癌高转移细胞系LoVo;siRNA介导的SIRT5基因沉默能显著抑制结肠癌癌细胞的迁移和侵袭能力;降低MMP2及MMP9的表达,并且使E-cadherin表达水平升高,N-cadherin、Vimentin表达水平降低。结论沉默SIRT5可以通过调节结肠癌细胞EMT,从而抑制结肠癌细胞的迁移和侵袭能力,可能为结肠癌治疗的潜在靶点。

Objective To investigate the effects of silent information regulator 5 （SIRTS） gene silencing on migra- tion and invasion of colonic carcinoma cells. Methods The expressions of SIRT5 were analyzed by reverse transcription- polymerase chain reaction （RT-PCR） and western blot in different colonic carcinoma cell lines and immortalized colonic epithelial cells. The expressions of matrix metalloproteinase （MMP） 2, MMP 9, epithelial-mesenchymal transition （EMT）, index protein （E-cadherin, N-cadherin, Vimentin） were tested by westdrn blot. Results SIRT5 was obviously over-expressed＇ in colonic carcinoma cell lines, especially in high metastastic potential colonic carcinoma cell line LoVo. Si- lencing of SIRT5 gene was significantly inhibited migratory and invasive capacities of colonic carcinoma cells （P〈0. 01）, decreased the expressions of MMP 2 and MMP 9, as well as up-regulated E-cadherin, down-regulated N-cadherin and Vimentin. Conclusion SIRT5 silencing gene suppress migratory and invasive capasities of colonic carcinoma by regulating epithelial-mesenchymal transition, and it may be potential target of cure for colonic carcinoma.