腺病毒Ela基因增强P16基因诱导SMMC-7721肝癌细胞的凋亡  

Adenovirus Ela gene enhances P16 gene-induced apoptosis of hepatocellular carcinoma SMMC-7721 cells

作　　者：胡还章[1,2] 王伟国 马炬明 苏长青[2] 江艺[1] 

机构地区：[1]南京军区福州总医院肝胆外科,福建福州350025 [2]第二军医大学东方肝胆外科医院病毒治疗研究室,上海200438 [3]南京军区117医院消化内科,浙江杭州310004

出　　处：《中国肿瘤生物治疗杂志》2011年第6期630-634,共5页Chinese Journal of Cancer Biotherapy

基　　金：国家自然科学基金资助项目(No.81071866,No.81172303)~~

摘　　要：目的:研究腺病毒Ela基因与P16抑癌基因协同对肝癌SMMC-7721细胞凋亡和增殖的影响,探索肿瘤基因治疗新模式。方法:构建Ela基因真核表达质粒pDC315-E1a和携带P16的重组病毒AdCMV-P16,RT-PCR和免疫荧光标记法检测pDC315-E1a质粒转染或AdCMV-P16病毒感染后SMMC-7721细胞中P16和Ela的表达。建立裸鼠SMMC-7721细胞移植瘤模型,pDC315-E1a和AdCMV-P16单独或联合治疗,观察其对移植瘤生长的抑制作用,免疫组化和TUNEL法分别检测移植瘤组织中P16、E1a蛋白的表达和移植瘤细胞的凋亡。结果:SMMC-7721细胞感染AdCMV-P16或转染pDC315-E1a后,P16、Ela mRNA和蛋白水平均呈阳性表达。与空白对照组相比,AdCMV-P16治疗组移植瘤细胞凋亡率为(14.3±2.5)%(P<0.01),抑瘤率为36.1%(P<0.01);pDC315-E1a治疗组抑瘤率为17.1%(P>0.05),移植瘤细胞凋亡率为(8.5±2.9)%(P<0.01);AdCMV-P16联合pDC315-E1a治疗组移植瘤细胞凋亡率为(27.3±6.3)%(P<0.01),抑瘤率达57.2%(P<0.01)。结论:腺病毒Ela基因能够增强P16基因对肝癌SMMC-7721细胞移植瘤的抑制作用,促进移植瘤细胞的凋亡,增强P16基因治疗的效果。Objective： To investigate the synergistic effect of anti-cancer P16 gene and adenovirus E1a gene on apoptosis and proliferation of hepatocellular carcinoma SMMC-7721 cells, and to explore the novel therapeutic strategy for tumor gene therapy. Methods： Eukaryotic expression plasmid pDC315-E1a and adenoviral vector AdCMV-P16 were constructed. The expression of P16 and E1a in SMMC-7721 cells after pDC315-E1a transfection or AdCMV-P16 infection was determined by RT-PCR and immunofluorescent labeling. SMMC-7721 cell transplanted tumors in nude mice was established. The effect of pDC315-El a and AdCMV-P16 alone or incombination on tumor growth was observed, and the expressions of P16 and El a in transplanted tumor tissues and apoptosis of transplanted tumor cells were determined by immunohistochemistry and TUNEL assay, respectively. Results： SMMC-7721 cells showed positive expression of both mRNA and protein levels of E1a and P16 after pDC315-E1a transfection or AdCMV-P16 infection, respectively. Compared with the control group, the apoptosis rate of transplanted tumor cells was （ 14.3 ＋ 2.5 ） % （ P 〈 0.01 ） and tumor inhibitory rate was 36. 1% （P〈0.01） in AdCMV-P16 therapy group; those in pDC315-E1a therapy group was （8.5 ±2.9）% （P〈0.01） and 17.1% （P 〉 0.05）; and in AdCMV-P16 combined pDC315-E1a therapy group was （27.3 ± 6.3）% （P 〈 0.01 ） and 57.2% （P 〈0.01 ）, respectively. Conclusion： Adenovirus E1a gene can increase P16-induced apoptosis and cell growth inhibition in SMMC-7721 cell transplanted tumors, and thus enhanee the efficaey of P16 gene therapy.

关 键 词：肝癌 P16基因 E1A基因 腺病毒 凋亡 

分 类 号：R735.7[医药卫生—肿瘤] R730.54[医药卫生—临床医学]