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Knockdown of survivin gene expression by RNAi induces apoptosis in human hepatocellular carcinoma cell line SMMC-7721

查看全文 作　　者：Sheng-[1]QuanCheng;Wen-[2]LiangWang;[2]WeiYan;Qing-[2]LongLi;[2]LiWang;Wen-[2]YongWang 高影响力作者 机构地区：[1]DepartmentofPediatrics,XijingHospital,FourthMilitaryMedicalUniversity,Xi'an710032,ShaanxiProvince,China;[2]DepartmentofPathology,FacultyofPreclinicalMedicine,FourthMilitaryMedicalUniversity,Xi'an710032,ShaanxiProvince,China高影响力机构 出　　处：《World Journal of Gastroenterology》索引2005年第11卷第5期,共4页高影响力期刊 摘　　要：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-SW), were constructed by ligating into the vector,pSupperssorNeo (pSuNeo) digested with restriction enzymes Xba I and Sa/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-avidinbiotin-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-SW, 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. 关 键 词：基因表达 RNAI 诱导作用 细胞凋亡 肝细胞癌 肿瘤细胞系统 SMMC-7721 抑制作用
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