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目的 观察单纯疱疹病毒 胸苷激酶 (HSV tk) /丙氧鸟苷 (GCV)基因治疗系统在体内外对人胰腺癌细胞的杀伤效应。方法 构建含HSV tk基因的重组逆转录病毒载体pDOR tk neo ,经病毒包装细胞PA31 7产生重组逆转录病毒 ,后者将HSV tk基因转入人胰腺癌细胞系PC 2细胞内 ,筛选出稳定表达HSV tk的细胞克隆PC 2 /tk。测定PC 2 /tk细胞在体外对GCV的敏感性及其旁观杀伤效应 ;PC 2 /tk细胞在裸鼠皮下成瘤 ,观察GCV的治疗作用。结果 GCV对PC 2 /tk细胞有明显的杀伤作用 ,且其作用呈现剂量和时间依赖性特点以及旁观杀伤效应 ,对母本细胞则无明显毒性。裸鼠腹腔注射GCV后对移植瘤有明显治疗作用。结论 表达HSV tk的人胰腺癌细胞在体内外均可被GCV有效杀伤。
Objective To observe the killing effect of HSV tk / GCV gene therapy system on human pancreatic cancer cells in vitro and in vivo. Methods The recombinant retroviral vector pDOR tk neo containing HSV tk gene was constructed and the recombinant retrovirus was generated by virus packaging cell PA31 7. The recombinant retroviral vector transfected HSV tk gene into human pancreatic cancer cell line PC 2 and screened out stable expression HSV tk cell clone PC 2 / tk. The sensitivity of PC 2 / tk cells to GCV and their bystander killing effect were determined in vitro. PC 2 / tk cells were subcutaneously injected into nude mice to observe the therapeutic effect of GCV. Results GCV had a significant killing effect on PC 2 / tk cells, and its effects were dose-and-time-dependent and bystander killing effect, but no obvious toxicity on maternal cells. Nude mice after intraperitoneal injection of GCV has obvious therapeutic effect on the xenografts. Conclusion Human pancreatic cancer cells expressing HSV tk can be effectively killed by GCV both in vitro and in vivo.