基因-病毒治疗系统CNHK300-小鼠内皮抑素对裸鼠胃癌的抗肿瘤作用  

Anti-tumor effect of gene-viral therapeutic system CNHK300-murine endostatin on nude mouse gastric cancer

作　　者：聂明明[1] 方国恩[1] 王星华[1] 苏长青[2] 钱其军[2] 

机构地区：[1]第二军医大学附属长海医院普通外科9B病区,上海200433 [2]第二军医大学附属东方肝胆医院病毒和基因治疗实验室,上海200433

出　　处：《中华胃肠外科杂志》2007年第6期565-569,共5页Chinese Journal of Gastrointestinal Surgery

摘　　要：目的探讨新构建的基因-病毒治疗系统CNHK300-小鼠内皮抑素murineendostatin（CNHK300-mE）对胃癌的抑制作用。方法通过胃癌SGC-7901细胞裸鼠皮下移植瘤模型观察该病毒治疗系统对胃癌生长和肿瘤血管生成的抑制作用。用电镜观察该病毒在肿瘤细胞中的复制情况及肿瘤细胞的超微结构改变。用酶联免疫吸附（ELISA）法检测mE基因在体内的表达。用免疫组织化学的方法检测腺病毒外壳蛋白六邻体（hexon）、增殖细胞核抗原（PCNA）及vWF因子相关抗原的表达情况。采用TUNEL法检测细胞凋亡。结果CNHK300-mE能在肿瘤细胞内复制,高表达mE,在第7天时,达到（2115±770）ng/ml（范围1745～3000ng/ml）;明显抑制胃癌皮下移植瘤的生长及瘤内的血管生成,并可引起胃癌细胞的凋亡[治疗组小鼠肿瘤细胞凋亡率（78.4±9.1）%,对照组仅（15.2±0.5）%,P〈0.01],抑制肿瘤细胞增殖[治疗组小鼠PCNA指数（55.0±1.4）%,对照组为（74.1±0.4）%,P〈0.05]。结论CNHK300-mE能在小鼠胃癌内增殖复制,高效表达mE基因,抑制胃癌生长。Objective To investigate the anti-tumor effect of a novel gene-viral therapeutic system CNHK300-murine endostatin （CNHK300-mE）on gastric cancer. Methods SGC-7901 gastric cancer cells （5×10^7 cells/mouse） were injected s.c. into the right flank of Bal b/c nude mice, grown to 4-5 mm to demonstrate tumor take, and 10^9 pfu/100 μl CNHK300-mE virus was injected into tumors. Tumor sizes were measured with calipers every other day. Serum samples were obtained by retro-orbital puncture and level of endostatin expression in serum was quantitated by ELISA. Fifteen days after treatment, all mice were sacrificed and tumors were excised for immunohistochemical staining of PCNA, hexon and vWF. Tumor cell apoptosis was detected by TUNEL method. Results From the 7th day post-treatment, the bearing tumors of mice treated with CNHK300-mE were significantly smaller than those of control group treated with PBS. Seven days after treatment, expression of endostatin was （2115±770） ng/ml, significantly higher than that of control group. Immunohistochemical staining indicated that hexon was expressed in treated tumor cells, and PCNA LI（label index）[（55.0±1.4）% vs control （74.1±0.4）%,P〈0.05], microvessel density （MVD） of CNHK300-mE treated tumors decreased significantly. Apoptosis obviously increased in tumor cells[（78.4±9.1）% vs control （15.2±0.5）%,P〈0.01]. Apoptosis bodies and crystal grid were found in tumor cell nuclear by electron microscope. Conclusions Gene-viral therapeutic system CNHK300-murine endostatin can replicate in gastric cancer cells. The mouse endostatin gene cloned into CNHK300-mE expresseds in high level. CNHK300-mE may induce tumor cells apoptosis, reduce the expression of PCNA and efficiently suppress gastric cancer growth through inhibiting tumor angiogenesis.

关 键 词：胃肿瘤 基因治疗 病毒 端粒酶 内皮抑素 

分 类 号：R735[医药卫生—肿瘤]