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首页> 外文期刊>World Journal of Gastroenterology >Knockdown of survivin gene expression by RNAi induces apoptosis in human hepatocellular carcinoma cell line SMMC-7721.
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Knockdown of survivin gene expression by RNAi induces apoptosis in human hepatocellular carcinoma cell line SMMC-7721.

机译：RNAi抑制survivin基因表达可诱导人肝癌细胞SMMC-7721凋亡。

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AIM: To investigate the survivin gene expression in human hepatocellular carcinoma cell line SMMC-7721 and the effects of survivin gene RNA interference (RNAi) on cell apoptosis and biological behaviors of SMMC-7721 cells. METHODS: Eukaryotic expression vector of survivin gene RNAi and recombinant plasmid pSuppressorNeo-survivin (pSuNeo-SVV), were constructed by ligating into the vector, pSuppressorNeo (pSuNeo) digested with restriction enzymes Xba I and Sal I and the designed double-chain RNAi primers. A cell model of SMMC-7721 after treatment with RNAi was prepared by transfecting SMMC-7721 cells with the lipofectin transfection method. Strept-avidin-biotin-complex (SABC) immunohistochemical staining and RT-PCR were used to detect survivin gene expressions in SMMC-7721 cells. Flow cytometry was used for the cell cycle analysis. Transmission electron microscopy was performed to determine whether RNAi induced cell apoptosis, and the method of measuring the cell growth curve was utilized to study the growth of SMMC-7721 cells before and after treatment with RNAi. RESULTS: The eukaryotic expression vector of survivin gene RNAi and pSuNeo-SVV, were constructed successfully. The expression level of survivin gene in SMMC-7721 cells was observed. After the treatment of RNAi, the expression of survivin gene in SMMC-7721 cells was almost absent, apoptosis index was increased by 15.6%, and the number of cells was decreased in G2/M phase and the cell growth was inhibited. CONCLUSION: RNAi can exert a knockdown of survivin gene expression in SMMC-7721 cells, and induce apoptosis and inhibit the growth of carcinoma cells.
机译：目的：探讨survivin基因在人肝癌细胞SMMC-7721中的表达以及survivin基因RNA干扰（RNAi）对SMMC-7721细胞凋亡及生物学行为的影响。方法：将Survivin基因RNAi的真核表达载体与重组质粒pSuppressorNeo-survivin（pSuNeo-SVV）连接，构建质粒后，将其分别用限制性内切酶Xba I和Sal I消化，并设计双链RNAi引物。 。用脂质体转染法转染SMMC-7721细胞，制备了RNAi处理后的SMMC-7721细胞模型。链霉亲和素-生物素复合物（SABC）免疫组化染色和RT-PCR用于检测SMMC-7721细胞中survivin基因的表达。流式细胞仪用于细胞周期分析。进行透射电子显微镜以确定RNAi是否诱导了细胞凋亡，并利用测量细胞生长曲线的方法研究了用RNAi处理之前和之后SMMC-7721细胞的生长。结果：成功构建了survivin基因RNAi和pSuNeo-SVV的真核表达载体。观察survivin基因在SMMC-7721细胞中的表达水平。 RNAi处理后，SMMC-7721细胞几乎不存在survivin基因表达，凋亡指数提高了15.6％，G2 / M期细胞数量减少，抑制了细胞生长。结论：RNAi可抑制SMMC-7721细胞中survivin基因的表达，并诱导其凋亡并抑制癌细胞的生长。

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