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目的 构建沉默信息调节因子(silent information regulaor,Sir)样蛋白1(Sirtuin 1,SIRT1)的特异性短发夹RNA(shRNA)慢病毒载体,获得敲低SIRT1的肝癌细胞系,以探讨其对肝癌细胞的增殖和耐药敏感性的作用.方法 设计针对SIRT1靶点特异性的干涉序列,连接到经HpaⅠ和XhoⅠ双酶切的pSicoR-GFP载体,慢病毒包装293T产生病毒,感染肝癌细胞,建立肝癌细胞SIRT1低表达的稳定株.利用实时定量PCR检测SIRT1的干涉效果;通过平板克隆形成实验、CCK8细胞增殖实验检测SIRT1被干涉后对肝癌细胞增殖能力的影响;通过细胞耐药敏感性实验及实时定量PCR检测耐药基因的表达,考察SIRT1干涉对肝癌细胞耐药敏感性的影响.结果 实时定量PCR实验证明该慢病毒干涉载体能显著抑制肝癌细胞中SIRT1的表达,SIRT1干涉可抑制肝癌细胞的增殖能力,下调耐药基因的表达,增强肝癌细胞的药物敏感性.结论 SIRT1抑制肝癌细胞耐药性.“,”Objective To build a short-hairpin RNA (shRNA)interference lentiviral vector of silent information regulator(sir)-like protein 1 (Sirtuin 1,SIRT1 )gene and to explore its effect on proliferation and drug resistance in hepatocellular carcinoma(HCC)cells.Methods The SIRI1 target specific oligonucleotide sequence was designed and connected to pSicoR-green fluorescent protein(GFP)vector digested by the HpaⅠ and XhoⅠ.Then,293T cells were packaged to produce the lentivirus and transfected with liver cancer cells.qRT-PCR assays further indicated that SIRT1 gene was stably knocked down in HCC cells.The effect on cell proliferation was detected by foci formation assay and CCK8 assay.In addition,drug resistance experiments and qRT-PCR were used to detect the effect on resistance of liver cancer cells when the SIRT1 gene was knocked down.Results The result of qRT-PCR demonstrated that the lentivirus shRNA vector of SIRT1 gene was constructed and could suppress the expression level of SIRT1.The proliferation and drug resistance genes of HCC cells were inhibited and drug sensitivity was enhanced.Conclusion SIRT1 gene represses the drug resistance of HCC cells.