携带IL-12的增殖腺病毒对鼻咽癌细胞的杀伤作用

目的：肿瘤选择性增殖腺病毒携带小鼠白细胞介素12（mIL-12)基因,增强对鼻咽癌细胞的杀伤作用。方法：用非增殖腺病毒Ad-LacZ测定腺病毒对鼻咽癌细胞CNE3和915的转染率,用E1B－55000u蛋白缺失的腺病毒dl1520和E1B－55000u蛋白缺失的腺病毒CNHK200－mlIL12分别转染培养的CNE3和915,并瘤内注射到裸鼠皮下的CNE3移植瘤。结果：Ad-LacZ病毒数量与细胞数量之比（MOI）为100时,转染率分别为63％和56％;MOI为10时,分别为12％和8％。 在体外不加入淋巴细胞的条件下,CNHK200－mIL和dl1520在MOI为100时都可在7d内完全杀灭鼻咽癌细胞CNE3和915。对裸鼠CNE3移植瘤,CNHK200－mil12治疗组疗效显著高于dl1520治疗组（P＜0．05） ,表明在选择性腺病毒中插入mIL12增强了其杀伤功能。结论：选择性增殖腺病毒携带mIL12后能够增强病毒对鼻咽癌细胞的杀伤作用,为鼻咽癌临床治疗探索一种新的基因病毒治疗方法。

Replicating adenovirus with mIL12 gene enhances the cytopathic effect of selective replicating adenovirus on nasopharyngeal carcinoma cells

Objective The selective replicate adenovirus was used to express mouse interleukin 12 (mIL12) in order to enhance the therapeutic effects on the nasopharyngeal carcinoma (NPC).Methods Replicate deficient adenovirus with LacZ gene(Ad LacZ) was used to assay the ability of adenovirus infected NPC, such as CNE3 and 915. CNHK200 mIL12 and dl1520 were trans infected into 915 and CNE3 and intratumorally injected into the CNE3 NPC of nude mice in subcutaneum to treat the tumor in vivo.Results When multiple of infect(MOI) was 100, Ad LacZ can infect 63% CNE3 and 56% 915 NPC cells. When MOI was 10, they were 12% and 8%, respectively. The results displayed that adenovirus can infect the NPC cells effectively. In vitro , when MOI was 100, the E1B 55 000 u gene depleted selective replicate adenovirus dl1520 and CNHK200 mIL12 can kill all of the NPC cells without lymphocytes in 7 days, respectively. There were no marked diffe rence between CNHK200 mIL12's cytopathic effect on NPC cells and that of dl1520. It proved cloning mIL12 into the selective replicate adenovirus had no effect on its cytopathic effect. Injecting into CNE3 NPC with CNHK200 mIL12 resulted in marked regression of the established tumors ascompared to injecting with dl1520 ( P <0.05). Conclusion This study proves CNHK200 mIL12 enhances the therapeutic effect on tumor by expressing mIL12. These data suggest that CNHK200 IL12, which augments the antitumor effect of replicate adenovirus, may be a powerful tool for treating NPC.