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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 effect of silent information regulator 5 (SIRT5) gene silencing on colon cancer cell migration and invasion. Methods The expression of SIRT5 in various colon cancer cell lines and immortalized colon epithelial cells was analyzed by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. Western blot was used to detect the expression of matrix metalloproteinase 2 (MMP2), matrix metalloproteinase 9 (MMP 9), epithelial mesenchymal transition (EMT) E-cadherin, N-cadherin, Vimentin]. Results Compared with normal human colon epithelial cells, the expression of SIRT5 was significantly up-regulated in colorectal cancer cell lines, especially LoVo, a highly metastatic human colorectal cancer cell line. SIRT5 siRNA-mediated silencing of SIRT5 gene significantly inhibited the migration and invasion of colon cancer cells Ability; reduce the expression of MMP2 and MMP9, and make E-cadherin expression level increased, N-cadherin, Vimentin expression levels decreased. Conclusion Silencing SIRT5 can inhibit the migration and invasion of colon cancer cells by regulating the EMT of colon cancer cells, which may be a potential target for the treatment of colon cancer.