摘要
目的将已构建完成的携带增强绿色荧光蛋白基因的TSLC1真核表达载体转染到HepG2细胞中,同时观察其对HepG2肝癌细胞的作用。方法脂质体Lipofectamine 2000介导重组载体转染到HepG2肝癌细胞中,经G418筛选建立稳定转染细胞株,分别采用RT-PCR、免疫组化法和westernblot法检测其表达;MTT检测细胞增殖活力,流式细胞仪分析细胞周期。结果与对照组比较,RT-PCR方法可见转染组细胞TSLC1mRNA表达明显增多,免疫组化和western-blot方法可见转染组细胞TSLC1蛋白表达明显增高;MTT检测发现转染组细胞生长缓慢,细胞周期检测发现转染组G0/G1期细胞显著增多,而S期及G2/M细胞明显降低。结论本研究成功建立了稳定转染TSLC1的HepG2细胞株,并证实TSLC1能够抑制细胞生长、阻滞细胞周期。
Objective To transfect TSLC1 gene of carrying enhanced green fluorescent protein of gene into HepG2 cell and wske TSLC1 express so as to investigate the mechanism TSLC1 in tumor suppression.Methods This research was divided into the control group and the transfected group. After the pReceiver-M29-TSLC1 and pReceiver-M29 were transfected into HepG2 hepatoma cell by liposome 2000, we had established stable transfected cell line using G418 screening and detected TSLC1 expression using RT-PCR, immunochemistry and Western blotting.The cell cycle was analyzed by flow cytometry （FCM）, and the cell activity was detected by M3T method. Results Comparing with the control group, the expression of TSLC1 mRNA and protein were significantly increasing in transfected group（ P 〈 0.05） ,the cell growth was slower,the percentage of G0/G1 phase was higher and the percentage of S and G2/ M phase was lower （ P 〈 0.05）. Conclusion We successfully construct the stable transfected HepG2 cell line with TSLC1 and verify that TSLC1 can inhibit HepG2 cell line growth and blockaging cell cycle.
出处
《中国实验诊断学》
北大核心
2010年第3期365-368,共4页
Chinese Journal of Laboratory Diagnosis
基金
吉林省发展与改革委员会资助课题[(2006)1550号]
