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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 the co-expression of HSV-TK and B7 gene in rat breast cancer cells. Methods Recombinant GcTKpB7SN, GcpB7SN and GcTKpSN retroviral vectors carrying TK and B7 genes were constructed respectively by gene recombination technique. The recombinant vectors were packaged and infected with virus supernatant to induce SHZ-88 rat breast cancer cells, and then G418 resistance Clone count, flow cytometry, in situ cell hybridization detection of B7, TK gene expression. Results Adenocarcinoma cells transfected with recombinant GcTKpB7SN and GcpB7SN genes showed the expression of B7 gene by flow cytometry. In situ cell hybridization showed that the expression of TK gene was observed in the transfected GcTKpB7SN and GcTKpSN breast cancer cells. Conclusion The retroviral vector-mediated exogenous suicide gene and costimulatory factor can be co-expressed in rat breast cancer cells.