目的 探讨沉默信息调节因子2相关酶3(Sirt3)蛋白对宫颈癌细胞脂质代谢的作用.方法 应用qRT-PCR和Western blotting技术检测Sirt3基因在宫颈癌SiHa和C33a细胞中的转录与蛋白本底表达情况,采用基因过表达和RNA干扰技术分别上调和下调子宫颈癌SiHa和C33a细胞中Sirt3的表达;Western blotting技术检测Sirt3蛋白水平变化.油红O染色技术检测转染Sirt3低表达和高表达慢病毒前后宫颈癌细胞脂质水平的变化情况.结果 与C33a细胞相比,SiHa细胞中Sirt3的mRNA与蛋白的表达水平均下降(P<0.05),故选用SiHa细胞转染Sirt3过表达慢病毒,C33a细胞转染Sirt3低表达慢病毒.SiHa细胞转染过表达慢病毒后,Sirt3蛋白表达水平明显增高(P<0.05),与正常对照组和阴性对照组比,转染Sirt3高表达慢病毒的细胞内脂质含量有所增加.C33 a细胞转染Sirt3低表达慢病毒后,Sirt3蛋白表达水平明显降低(P<0.05),与正常对照组和阴性对照组比,转染Sirt3低表达慢病毒的细胞内脂质含量降低.结论 Sirt3蛋白的表达参与宫颈癌细胞内的脂质代谢,具体调控机制尚待研究.%:Objective To explore the effect of Sirt3 protein expressions on Lipid Metabolism of Cervical Cancer Cells.Methods mRNA and the protein of Sirt3 were detected in normal cervical cancer SiHa and C33a cell by qRT-PCR and Western blotting.The Sirt3 expression was down-regulated in C33a cell by RNA interferencing technology,while the Sirt3 gene was up-regulated in SiHa cell through gene overex-pression technology.After transfected lentiviral vector,the Sirt3 protein expression was detected by West-ern blotting.Oil red O staining was used to evaluate the changing of lipid levels in cervical cancer cells in-fected lentiviral vector and the control transfected cells and normal cervical cancer cells.Results The mR-NA and protein level of Sirt3 in SiHa cell were significantly lower than that in C33a cell (P<0.05),so Si-Ha cell was transfected with SIRT3 overexpression lentiviru,and C33a cell was transfected with SIRT3 siRNA lentiviru.The expression of Sirt3 protein and the content of intracellular lipid were significantly in-creased in SiHa cells after transfected with overexpression lentiviru compared to the control and normal cell (P <0.01).While the level of Sirt3 protein was significantly decreased (P <0.01)and reduced the content of intracellular lipid in C33a cell transfected with siRNA lentivirus than the control and normal cell.Conclusion The expression of Sirt3 protein participates in lipid metabolism in cervical cancer cells, but the specific regulation mechanism remains to be investigated.
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