摘要
目的 研究含单纯疱疹病毒Ⅱ型胸苷激酶基因 (HSV ⅡTK)对人原发性肝细胞癌SMMC 772 1的治疗作用。方法 用已构建的真核表达载体 pcDNA3/TK ,通过阳离子脂质体Lipo fectin介导 ,体外转染人原发性肝细胞癌SMMC 772 1。加入GCV ,细胞培养。用RT PCR ,光镜 ,MTT ,流式细胞仪等方法检测HSV ⅡTK在SMMC 772 1细胞中的表达及其对该细胞的杀伤作用。结果 实验组细胞通过RT PCR获得与目的基因 (HSV ⅡTK)大小一致 (10 0 0bp)的特异性条带 ,而对照组细胞则无任何条带出现 ;MTT法检测显示 ,实验组的OD值明显低于对照组 ,且有显著意义(P <0 0 1) ;流式细胞仪检测显示 ,实验组可发现明显的凋亡峰 ,而对照组则无凋亡峰出现。结论 通过阳离子脂质体Lipofectin介导 ,HSV ⅡTK在SMMC 772 1细胞中能够获得表达 ,其表达产物结合GCV可明显抑制人原发性肝细胞癌细胞的增殖。
Objective To investigate the therapeutic effects of thymidine kinase (TK) gene of herpes simplex virus Ⅱ (HSV-Ⅱ) on human primary hepatocellular carcinoma (HPHCC). Methods The constructed eukaryon expression vector pcDNA3/TK was transferred into HPHCC cell line SMMC-7721 in vitro by cation liposome lipofectin. RT-PCR, MTT and FCM were used to detect expression of HSV-Ⅱ TK in the SMMC-7721 cells and its inhibitory effects on the cells. Results A 1000bp specific fragment that is the same as HSV-Ⅱ TK was obtained through RT-PCR. It was found that the HSV-Ⅱ TK gene was expressed in the SMMC-7721 cells. MTT showed that the HSV-Ⅱ TK/GCV system could efficiently inhibit the growth of SMMC-7721 cells. The peak of cell apoptosis was seen experimental group but not in the control by FCM. Conclusions After being transferred by cation liposome lipofectin, HSV-Ⅱ TK gene can be expressed in SMMC-7721 cells. The HSV-Ⅱ TK/GCV system can efficiently inhibit the growth of HPHCC cells.
出处
《中华肝胆外科杂志》
CAS
CSCD
2003年第5期306-308,共3页
Chinese Journal of Hepatobiliary Surgery
