重组Ad-CRM8-survivin-EHV4 TK腺病毒的构建及对肝癌移植瘤的抑制作用

Construction of adenovirus carrying HS-CRM8,surviving promoter and EHV4 TK and its inhibitory effect on the xenograft tumor of HCC in nude mice

目的:构建含有肝癌特异性的HS-CRM8增强子、survivin启动子及EHV4 TK自杀基因的腺病毒,探讨该重组病毒对裸鼠肝癌移植瘤的抑制作用。方法:构建重组Ad-CRM8-survivin-EHV4 TK腺病毒,获取重组腺病毒。构建裸鼠肝癌皮下移植瘤模型,给予重组腺病毒结合更昔洛韦(GCV)治疗,2周后观察肿瘤大小。对肿瘤组织切片使用HE染色,观察瘤内细胞坏死情况。结果:裸鼠肝癌移植瘤经腺病毒治疗后,EHV4 TK和GCV+EHV4 TK组肿瘤体积分别为(108.5±18.3)mm3和(37.6±8.7)mm3。GCV+EHV4 TK组与EHV4 TK相比,裸鼠的肿瘤重量减少,生存率提高。HE染色结果显示,GCV+EHV4 TK组肿瘤内出现大量细胞的坏死,而EHV4 TK组肿瘤内仅有少量细胞的坏死。结论:重组肝脏特异性腺病毒Ad-CRM8-survivin-EHV4 TK结合GCV能够有效地在体内促进肝癌细胞的坏死从而抑制裸鼠移植瘤的生长,对肝癌的治疗有良好的效果。

Objective：To evaluate the inhibitory effect of adenovirus-mediated HS-CRM8-survivin-EHV4 TK suicide gene on the xenograft tumor of hepatocellular carcinoma（HCC）in nude mice. Methods：A liver-specific adenovirus vector,Ad-CRM8-survivinEHV4 TK（EHV4 TK）,was constructed by molecular cloning and recombination. In nude mice bearing human hepatocellular carcinoma,the recombinant adenovirus was injected directly into the tumor followed by intraperitoneal injection of the prodrugs GCV and the inhibition effect on tumor xenografts was observed. The tumor section was stained with hematoxylin and eosin. Results：After adenovirus treatment, the volume of the tumors in EHV4 TK and GCV＋EHV4 TK groups were（108.5 ± 18.3）and（37.6 ± 8.7）mm^3.GCV＋EHV4 TK represented much stronger anti-tumor effect on tumor growth than EHV4 TK only. GCV＋EHV4 TK also showed much more tumor necrosis than EHV4 TK only. Conclusion：The adenovirus-mediated HS-CRM8-survivin-EHV4 TK suicide gene combined with GCV treatment may provide us a more effective,specific and convenient gene therapy method.