逆转录病毒介导HSV-TK基因治疗胰腺癌——实验研究

目的:探讨应用逆转录病毒载体介导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SV_(40) 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:Southern-blot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。体外试验证实,HSV-TK阳性8988细胞对ACV的敏感性较母细胞明显为高;裸鼠移植瘤试验证实,腹腔注射ACV有明显阻止移植瘤的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。

Experimental Studies on Pancreatic Cancer Gene Therapy with HSV-TK Gene Mediated by Retrovirus Vector

Background/Aims: To observe the value of HSV-TK gene mediated by retroviral vector in the treatment of human pancreatic cancer cell line-8988. Methods: HSV-tk gene was constructed into the site following SV40 promoter which is second to 5'LTR and neomycin phosphotransferase in the retroviral vector pMNSM which is able to integrate into host chromosomes. The recombinant plasmid pMNS-TK-M was transfected into the retroviral packaging cell PA317. Finally HSV-TK gene was transferred into human pancreatic cancer cell line-8988 by recombinant virus. Results: HSV-TK was integrated into host cell and .expressed by Southern blot test and drug-sensitivity test. In vitro study it was shown that ACV sensitive level of 8988/TK was obviously higher than that of its parent cell in culture. ACV showed a significant killing effect on the cells which were transducted with TK gene. In vivo study it was shown that intraperitoneal injection of ACV might postpone the formation of implanted tumors and was effective in nude mice. Conclusion: This study demonstrates that HSV-TK/ ACV can be used in vivo to treat pancreatic cancer and this method may be used as an innovative way for pancreatic cancer gene therapy in the future.