CAJ | 학술논문

目的构建双调控溶瘤腺病毒,携带小鼠内皮抑素基因(mE),研究其对裸鼠肝癌移植瘤的抗瘤活性.方法以人端粒酶逆转录酶启动子(hTERT)和缺氧调控元件序列(HRE)调控腺病毒E1a和E1b基因,基因组插入mE基因,构建双调控溶瘤腺病毒CNHK500-mE；在裸鼠模型中观察CNHK500-mE对肝癌移植瘤模型的疗效.结果 CNHK500能够在hTERT阳性的肝癌细胞中增殖[24 h:(16.67±4.04)％；48 h:(65.33 ±7.02)％;P＜0.01],并介导mE高效表达；与空白对照组( 1895.80±323.37) mm3比较,CNHK500-mE和Ad-mE的抑瘤率分别为50.95％[(929.80±211.10) mm3,P＜0.01]和29.99％[(1327.23 ±319.36) mm3,p＜0.05]；CNHK500-mE对癌组织间质血管的抑制作用明显强于Ad -mE(P ＜0.05).结论将mE与溶瘤腺病毒结合,发挥病毒增殖的溶瘤作用和基因产物的血管抑制作用,表现出明显的协同抗瘤作用.
목적 구건쌍조공용류선병독,휴대소서내피억소기인(mE),연구기대라서간암이식류적항류활성.방법 이인단립매역전록매계동자(hTERT)화결양조공원건서렬(HRE)조공선병독E1a화E1b기인,기인조삽입mE기인,구건쌍조공용류선병독CNHK500-mE；재라서모형중관찰CNHK500-mE대간암이식류모형적료효.결과 CNHK500능구재hTERT양성적간암세포중증식[24 h:(16.67±4.04)％；48 h:(65.33 ±7.02)％;P＜0.01],병개도mE고효표체；여공백대조조( 1895.80±323.37) mm3비교,CNHK500-mE화Ad-mE적억류솔분별위50.95％[(929.80±211.10) mm3,P＜0.01]화29.99％[(1327.23 ±319.36) mm3,p＜0.05]；CNHK500-mE대암조직간질혈관적억제작용명현강우Ad -mE(P ＜0.05).결론 장mE여용류선병독결합,발휘병독증식적용류작용화기인산물적혈관억제작용,표현출명현적협동항류작용.
Objective Dual-regulated oncolytic adenovirus carrying mouse endostatin gene (mE) was constructed,and its antitumor activity was examined in hepatocellular carcinoma (HCC) xenografts in nude mice.Methods Human telomerase reverse transcriptase promoter (hTERT) and hypoxia regulatory element (HRE) were used to control adenoviral Ela and E1b genes.The mouse endostatin gene was inserted into the viral genome to generate the dual-regulated oncolytic adenovirus CNHKS00-mE.Its antitumor activity was observed in HCC models in nude mice.Results CNHK500 could replicate in hTERT-positive HCC cells [24 h:( 16.67 ± 4.04) ％ ; 48 h:(65.33 ± 7.02) ％ ;P ＜ 0.01],and efficiently mediate endostatin expression; As compared with the control group ( 1895.80 ± 323.37) mm3,CNHK500-mE and Ad-mE showed antitumor efficacy on the growth of HCC xenografts,with the tumor inhibition rate of 50.95％ [(929.80±211.10) mm3,P＜0.01] and 29.99％ [(1327.23±319.36) mm3,P＜0.05],and the effect of CNHK500-mE was stronger than that of Ad-mE (P ＜0.01 ).Pathological examinations of xenografi tumors showed that CNHK500-mE proliferated in cancer cells and expressed high levels of endostatin,and the inhibitory effect of CHNK500-mE on tumor microvessels was also stronger than that of AdmE ( P ＜ 0.05 ).Injection of CHNK500-mE resulted in large area of tumor necrosis.Conclusion Combination of endostatin gene and dual-regulated oncolytic adenovirus exerts not only the oncolytic effect but also the tumor microvessel inhibition,finally produces the synergistic effect and enhances the antitumor efficacy.