逆转录病毒介导单纯疱疹病毒-胸苷激酶基因治疗胰腺癌的实验研究

Experimental study on pancreatic cancer gene therapy with HSV-TK gene mediated by retroviral vector

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 study the value of HSV TK gene mediated by retrovirus vector in the treatment of human pancreatic cancer cell line 8988. Methods HSV TK gene was constructed into the site following SV40 promoter which was second to 5′LTR and neomycin phosphotransferase in retroviral vector pMNSM. This enabled the above gene integrating into host chromesomes. The recombinant plasmid pMNS TK M was transfected into the retrovirus packaging cell PA317. Finally HSV TK gene was transferred into the pancreatic cancer cell line 8988 by recombinant virus. Results HSV TK had been stably integrated in host cell and expressed by Southern Blot and drug sensitivity tests. This in vitro study showed that ACV sensitivity level of 8988/TK + was higher than that of the cultured parent cell. ACV showed a significant killing effect on the cells which were transducted with TK gene. The in vivo study in nude mice showed that intraperitoneal injection of ACV might postpone the formation of the implanted tumors and had a treatment effect on the tumors. Conclusion This study demonstrates that HSV TK/ACV may be used in vivo to treat pancreatic cancer and may become a potential therapeutic modality in the future.