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目的 探讨应用逆转录病毒载体介导单纯疱疹病毒 胸苷激酶 (herpessimplexvirustypeIthymidinekinasegene,HSV TK)基因治疗实验性人胰腺癌细胞系 8988的价值。 方法 HSV TK被定向克隆入逆转录病毒载体 pMNSM的SV4 0 下游。重组逆转录病毒载体 pMNS TK M转染至逆转录病毒包装细胞PA317细胞 ,产生的重组病毒将HSV TK转入人胰腺癌细胞系 8988细胞内。结果 Southernblot试验及药敏试验均证实HSV TK基因已整合至细胞DNA中并完全表达。体外试验表明 ,HSV TK阳性 8988细胞对ACV的敏感性较母本细胞明显高 ;裸鼠移植瘤试验证实 ,腹腔注射ACV 10 0mg/kg有明显阻止移植瘤形成以及对移植瘤的治疗作用。结论 HSV TK/ACV有体内治疗胰腺癌的作用 ,可作为胰腺癌基因治疗的潜在方法之一。
Objective To investigate the value of retrovirus vector-mediated herpes simplex virus thymidine kinase gene (HSV TK) gene therapy in human pancreatic cancer cell line 8988. Methods HSV TK was directionally cloned into SV40 downstream of retroviral vector pMNSM. The recombinant retrovirus vector pMNS TK M was transfected into the retrovirus packaging cell PA317 cells, and the resulting recombinant virus transfected HSV TK into human pancreatic cancer cell line 8988. Results Both Southern blot and susceptibility tests confirmed that the HSV TK gene was integrated into the cell DNA and expressed completely. In vitro experiments showed that HSV TK positive 8988 cells were more sensitive to ACV than maternal cells; transplanted tumor in nude mice confirmed that intraperitoneal injection of ACV10 0mg / kg significantly prevented the formation of xenografts and the therapeutic effect on xenografts. Conclusions The role of HSV TK / ACV in the treatment of pancreatic cancer in vivo may be one of the potential methods for the gene therapy of pancreatic cancer.