应用基因转染方法调节HSP70在人胃癌细胞株SGC7901中的表达

Regulation of HSP70 expression in human gastric cancer cell line SGC7901 by gene transfection

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.