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目的 观察羟甲基无环鸟苷 (GCV)、CTL对逆转录病毒介导的转染单纯疱疹病毒胸苷激酶 (HSV TK)和共刺激分子 (B7)双基因的大鼠乳腺癌细胞的体外杀伤作用。方法应用基因重组技术 ,构建逆转录病毒载体GcTKpSN、GcpB7SN和GcTKpB7SN。以重组逆转录病毒感染细胞的培养上清液 ,感染大鼠乳腺癌细胞SHZ 88。经G4 18筛选阳性克隆并扩增后 ,给予GCV进行敏感性实验。将从正常SD大鼠脾细胞悬液中分离的T细胞 (CTL) ,与转染重组基因的乳腺癌细胞按不同比例共育后 ,进行细胞杀伤实验。结果 转染与未转染GcTKpB7SN基因的乳腺癌细胞相比较 ,前者对GCV的敏感性明显增强 (P <0 .0 1)。T细胞对经GcTKpB7SN基因修饰的乳腺癌细胞的杀伤作用 ,明显强于未转染Gc TKpB7SN基因的乳腺癌细胞 (P <0 .0 1)。结论 GCV对转染GcTKpB7SN基因的大鼠乳腺癌细胞具有体外杀伤作用。经GcTKpB7SN基因修饰的乳腺癌细胞 ,在体外能被T细胞杀伤。
Objective To observe the effects of hydroxymethyl acyclovir (GCV) and CTL on the expression of retroviral-mediated rat breast cancer cells transfected with herpes simplex virus thymidine kinase (HSV TK) and costimulatory molecule (B7) Killing effect. Methods Retroviral vectors GcTKpSN, GcpB7SN and GcTKpB7SN were constructed by gene recombination technique. The recombinant plasmid was transfected into the culture supernatant of retroviral infected cells to infect rat breast cancer cells SHZ 88. After positive clones were screened by G418 and amplified, GCV was given for sensitivity experiments. T cells (CTLs) isolated from the spleen cell suspensions of normal SD rats were co-cultured with the breast cancer cells transfected with recombinant genes at different ratios to perform cell killing experiments. Results Compared with non-transfected GcTKpB7SN gene, the former showed a significantly increased sensitivity to GCV (P <0.01). The cytotoxicity of T cells to GcTKpB7SN gene-modified breast cancer cells was significantly stronger than that of the non-transfected Gc TKpB7SN gene (P <0.01). Conclusion GCV can in vitro kill rat breast cancer cells transfected with GcTKpB7SN gene. The GcTKpB7SN gene modified breast cancer cells, in vitro T cells can be killed.