摘要
目的:探讨survivin反义RNA/HSP70双基因转染对肝癌细胞系HepG2的影响。方法:将已成功获得的双基因载体(pIRES2-EGFP-survivin反义RNA/HSP70)用脂质体介导转染肝癌细胞系HepG2,转染72小时,收集各组细胞,利用倒置荧光显微镜观察转染后的细胞,通过RT-PCR、western-blot检测转染后细胞中survivinmRNA、survivin蛋白及HSP70蛋白表达的变化。结果:转染双基因载体与转染空载体的肝癌细胞系HepG2于倒置荧光显微镜下可见绿色荧光;转染双基因载体的HepG2细胞与转染空载体的HepG2细胞和未转染的HepG2细胞相比,survivin mRNA和survivin蛋白表达水平降低,而HSP70蛋白表达升高。结论:Sur-vivin反义RNA与HSP70双基因转染成功;survivin反义RNA能干扰HepG2内源survivin的表达,HSP70能上调HepG2细胞HSP70蛋白的表达。
Objective：To explore the effects of survivin antisense RNA and HSP7 doublegene expression vector on hepatoma carcinoma cell line HepG2. Methods： Hepatoma carcinoma cell line HepG2 cells was transfected by lposome with double - gene vectors （ pIRES2 - EGFP - survivin antisense RNA /HSP7 ） obtained previously. The cells of all groups were collected when the cells had been transfected for 72 hours and were observed with inverted fluorescent microscope. We detected the changes of the expression of survivin mRNA, survivin protein and HSP70 prorein in the cells with RT - PCR and western - blot before and after the cells were transfected. Results： Green fluorescence in HepG2 cells transfected with the double -gene expression vector and transfected with the empty vector was observed under the inverted fluorescent microscope. The expression level of survivin mRNA and survivin protein reduced and the expression level of HSP70 protein enhanced in the HepG2 cells transfeeted with the double - gene expression vector, compared with the HepG2 cells transfected with the empty vector and the HepG2 cells which were not transfected. Conclusion： The double -gene expression vector was transfected successfully. Survivin antisense RNA can interfere with the expression of endogenous survivin in HepG2 cells. HSP70 can increase the expression of HSP70 protein in HepG2 ceils.
出处
《现代肿瘤医学》
CAS
2008年第5期694-696,共3页
Journal of Modern Oncology
基金
内蒙古医学院重大课题资助项目(NY2003ZD002)
教育部"春晖计划"资助项目(Z2005-2-01007)
