双基因导入在大鼠乳腺癌细胞内的共表达

COEXPRESSION OF HERPES SIMPLEX VIRUS THYMIDINE KINASE AND COSTIMULATORY MOLECULE GENES IN RAT NLAMMARY ADENOCARCINMA

目的观察携带单纯疱疹病毒胸苷激酶(HSV—TK)、共刺激因子(B7)基因联合在大鼠乳腺癌细胞的共表达。方法应用基因重组技术,分别构建携带含TK、B7基因的重组GcTKpB7SN,GcpB7SN、GcTKpSN逆转录病毒载体,重组载体经包装以病毒上清液感染SHZ-88大鼠乳腺癌细胞,而后进行G418抗性克隆计数,流式细胞、原位细胞杂交检测B7、TK基因表达。结果转染重组GcTKpB7SN、GcpB7SN基因的腺癌细胞经流式细胞仪检测表明有B7基因表达;原位细胞杂交对转染重组GcTKpB7SN、GcTKpSN基因乳腺癌细胞能观察到TK基因表达。结论经逆转录病毒载体介导外源性自杀基因和共刺激因子能在大鼠乳腺癌细胞内共表达。

Objective To observe coexpression of herpes simplex vires thymidine kinase(HSV-TK)and costimulatory tmolecule(B7)genes in rat mammary adenocarcinolma.Method The GcTKpSN、GepB7SN and GeTKpB7SN constructs were generated and insert-ed into retroviral vectors.Rat mammary adenocarcinoma cells SHZ-88 were trensduced with supernatant of recombinant retroviral con-stracts,cultured and selected with G418 resistance.The expression of B7 and TK genes was detected with flowcytometer and hybridiza-tion in situ respectively.Result mammary adenocarcinoma cells transduced with recombinant GcpE7SN and GeTKpB7SN genes had TKgene expression detected with flow cytometer;TK gane expression could be detected with hybridization in situ in cells transduced withGepTKSN and GeTKpB7SN genes.Conclusion exogenous suicide gene and costimuiatory molecule genes could be exressed in rat mamma-ry adenocarcinoma with retrovirus as transducing vector.