利用HSV-TK基因治疗肝癌的离体研究

GENE THERAPY FOR HEPATIC CANCER WITH HSVTK/GCV SYSTEM IN VITRO

利用ＤＮＡ重组技术，将ＨＳＶ－ＴＫ基因克隆至逆转录病毒载体（ＸＭ－６／ＴＫ）。经ＰＡ３１７细胞包装后，转染人肝癌细胞ＨｅｐＧ２。结果显示，ＸＭ－６／ＴＫ转染ＨｅｐＧ２细胞与野生型相比，其生长特点和细胞形态未有改变。给予抗病毒药物－环氧鸟苷（Ｇａｎｃｉｃｌｏｖｉｒ，ＧＣＶ）后，转染细胞生长受到严重抑制，细胞数量明显降低。细胞学检查证实，ＧＣＶ处理后，转染细胞的死亡率显著增加。本结果提示。

 retroviral vector XM6/TK containing HSVTK gene has been constructed using DNA recombinant technique.The retroviral recombinant was then transduced into PA317 packaging cell line by calcium coprecipitation.The supernatant of the packaging cells was used to transfect hepatoma cells HepG2. The results showed that the growth characteristics and cell morphology of the TK transfected hepatoma cells appeared to be little difference from those of wildtype HepG2 cells. It had been observed that the induction of GCV caused the obvious growth inhibition of the transfected cells, Meanwhile, the cell counts of the transfected cells decreased rapidly. The morphological examination proved that reduction of cell counts in transfected cells treated by GCV was caused by increased mortality. The results implied that the availability of HSVTK gene therapy might be one of the prospective alternatives for treatment of hepatocellular carcinoma.