摘要
目的 为探讨 HSP70 在人胃癌中过度表达的意义, 调节HSP70 在人胃癌细胞系SGC7901 中的表达.方法 应用脂质体介导的基因转染方法,将表达针对HSP70的反义RNA 表达载体及HSP70 表达载体转入SGC7901 细胞,通过hygro m ycine 抗性筛选, 挑选阳性克隆;对阳性克隆从RNA 水平( 点杂交及RNase 保护试验) 及蛋白质水平( Westernblot 及免疫组化) 进行研究.结果 HSP70 在转染细胞中的表达得到了调节. 为进一步研究HSP70 对胃癌形成与发展的作用创造了条件.结论 经过HSP 反义或正义RNA 转染的SGC7901 细胞具有不同的HSP70 表达水平可用于胃癌发生机制的研究.
AIM To regulate the expression of HSP70 in human gastric cancer cell line SGC7901 for clarifying the role of HSP70 in carcinogenesis of human gastric cancer. METHODS An antisense RNA to HSP70 expressing vector pBBS212/*!as hsp70 and a sense RNA to HSP70 expressing vector pBBS212/*!HSP70 were constructed and transfected into human gastric cancer cell line SGC7901 via lipofectamine mediated transfection. The clones of transfected cells were selected with hygromycine. The expression of HSP70 in the transfected cells was evaluated at mRNA level (dot blotting and RNase protection assay) and protein level (Western blotting and immunohistochemical staining). RESULTS The expression of HSP70 was successfully regulated in different transfected cells, up regulated in pBBS212/*!hsp70 transfected group and down regulated in pBBS212/*!as hsp70 transfected group. CONCLUSION The cells expressing different levels of HSP70 can be used for further studies concerning the role of HSP70 in carcinogenesis of human gastric cancer.
出处
《世界华人消化杂志》
CAS
1999年第9期773-776,共4页
World Chinese Journal of Digestology
关键词
热休克蛋白
胃肿瘤
基因转染
heat shock protein
stomach neoplasms
transfection