新型血管生成抑制人内皮细胞抑制素血管内皮细胞生长抑制因子_(151)治疗胃癌的实验研究  

The experimental studies of a novel hENDO-VEGI_ (151) fusion gene therapy on gastric carcinoma in nude mice mediated by adenovirus vector

作　　者：李喆[1] 方国恩[1] 闻兆章[1] 华积德[1] 曹贵松[1] 毕建威[1] 戚中田[2] 潘卫[2] 

机构地区：[1]第二军医大学附属长海医院普通外科,上海200433 [2]第二军医大学微生物学教研室

出　　处：《中华实验外科杂志》2005年第2期230-234,F003,共6页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目 (39970 81 9)

摘　　要：目的　研究重组腺病毒表达的人内皮细胞抑制素 血管内皮细胞生长抑制因子151(hENDO VEGI151)融合蛋白对胃癌的抑制作用。方法　构建携带hENDO VEGI151融合基因的重组腺病毒载体 ,脂质体介导法包装重组腺病毒AdIL 3 /hENDO VEGI151。体外检测融合基因的表达及融合蛋白的生物学活性。应用鸡胚绒毛尿囊膜 (CAM )模型及荷人胃癌裸鼠模型 ,进一步观察融合蛋白对活体血管生成的影响和融合基因治疗活体胃癌的疗效。结果　用TCID50 法测定携带融合基因的重组腺病毒滴度为 4.2× 10 11TCID50 /ml;用聚合酶链反应 (PCR)、逆转录 (RT) PCR和免疫组织化学方法证实融合基因可被转导入SGC 790 1细胞内并稳定高效地转录和表达 ;Westernblot显示融合蛋白可被分泌到胞外发挥作用 ;融合蛋白可强烈抑制ECV 3 0 4细胞生长及鸡胚绒毛尿囊膜新生血管形成。AdIL 3 /hENDO VEGI151治疗可强烈抑制裸鼠体内种植瘤生长 ,明显下调肿瘤微血管密度 ,促进胃癌细胞凋亡。结论　hENDO VEGI151是一条新型强效的肿瘤血管生成抑制基因 ,其表达产物可能通过作用于肿瘤新生血管形成的不同环节强烈抑制新血管生成和肿瘤生长 ,值得进一步研究。Objective To study the inhibitory effects of recombinant adenoviruses carrying the IL-3/hENDO-VEGI_ 151 fusion gene therapy on gastric carcinoma in nude mice.Methods Replication-defective recombinant adenovirus vector containing IL-3/hENDO-VEGI_ 151 gene was constructed.The recombinant adenovirus were packaged and used to infect SGC-7901 cells.The expression of the fusion gene was detected.CAM vascular inhibition test was used to evaluate the effect of recombinant adenovirus Ad IL-3/hENDO-VEGI_ 151.The recombinant adenovirus were injected into the established gastric carcinoma in nude mice.The expression of fusion gene and intratumoral microvessel density (MVD),proliferating cell nuclear antigen label index (PCNALI) and apoptotic index (AI) were evaluated respectively after the mice were sacrificed.Results Titer of the recombinant adenovirus was about 4.2×10 11TCID_ 50/ml.The fusion gene could be expressed in the gastric carcinoma SGC-7901 cell.The fusion gene could inhibit angiogenesis in CAM.Tumor growth was inhibited in nude mice subject to injection of the recombinant adenoviruses when compared to control mice treated with AdLacZ.Immunohistochemistry revealed tumor cell could be stained by Anti-endostatin,indicating that the chimeric gene could be expressed in the tumor.The MVD was also decreased significantly,while AIwas increased in the treated mice.Conclusion The recombinant adenovirus containing fusion gene of IL-3/hENDO-VEGI_ 151 can express the fuion protein in gastric carcinoma in nude mice and inhibit the tumor growthby a synergistic antiangiogenic mechanism.

关 键 词：治疗 融合基因 胃癌 人内皮细胞 融合蛋白 血管内皮细胞 重组腺病毒 VEGI 生长抑制因子 血管生成抑制 

分 类 号：R735.2[医药卫生—肿瘤] Q78[医药卫生—临床医学]