抗血管内皮生长因子发夹状核酶基因抑制卵巢癌移植瘤生长的实验研究  

Experimental study of anti-vascular endothelial growth factor hairpin ribozyme gene inhibiting growth of xenografted tumor of ovarian cancer cells

作　　者：严瑞兰[1] 辛晓燕[1] 金明[2] 惠宏襄[2] 王健[1] 王德堂[1] 

机构地区：[1]第四军医大学西京医院妇产科,西安710032 [2]第四军医大学西京医院生物化学与分子生物学教研室,西安710032

出　　处：《中华妇产科杂志》2002年第11期683-686,共4页Chinese Journal of Obstetrics and Gynecology

基　　金：国家自然科学基金资助项目 (3970 0 14 7)

摘　　要：目的　采用抗血管内皮生长因子 (VEGF)发夹状核酶基因 ,阻断卵巢癌中VEGF的自分泌和 (或 )旁分泌通路 ,以检测抗VEGF发夹状核酶基因对卵巢癌细胞VEGF表达及其肿瘤生长的影响。方法　采用脂质体介导的方法 ,将自行设计和构建的抗VEGF发夹状核酶基因真核表达载体转染人卵巢癌细胞SKOV3,采用氨基糖甙庆大霉素 (G418)筛选获得阳性克隆 ;RNA斑点杂交检测核酶基因在卵巢癌细胞中的表达 ;逆转录 聚合酶链反应 (RT PCR)技术检测转染前后卵巢癌细胞中VEGF的表达 ;透射电子显微镜观察卵巢癌细胞超微结构改变 ;观察裸鼠皮下成瘤实验检测转染前后细胞的致瘤能力的变化。结果　转染核酶基因后的卵巢癌细胞VEGFmRNA表达明显降低 ;透射电子显微镜观察出现凋亡细胞 ;裸鼠致瘤能力减弱 ,肿瘤组织中VEGF表达及血管形成减少。结论　抗VEGF发夹状核酶基因可显著抑制卵巢癌细胞VEGF表达 ,通过减少血管形成抑制肿瘤生长 ,为进一步开展肿瘤血管靶向基因治疗 。Objective We propose to block the autocrine and (or) paracrine pathway of vascular endothelial growth factor(VEGF)in ovarian cancer by using anti VEGF hairpin ribozyme gene to see whether the growth of tumor cells could be inhibited and to further exploit its mechanisms Methods Anti VEGF hairpin ribozyme gene eukaryotic expression vector was introduced into ovarian cancer cells SKOV 3 by using lipofectin mediation and positive clones were screened by genticin (G418); ribozyme (RZ) expression was confirmed by RNA dot blot; the VEGF expression of SKOV 3 cells before or after transfection were detected by reverse transcription polymerase chain reaction (RT PCR) method, transmission electron microscopy demonstrated the morphologic changes of ribozyme generating SKOV 3 cells; the growth of transfected cells in nude mice were detected Results VEGF expression was decreased distinctly in SKOV 3 cells transfected by RZ (SKOV 3 RZ) cells Apoptosis cells were observed; the formation rate and growth speed of xenografted tumor slowed down Conclusions Anti VEGF hairpin ribozyme gene can inhibit the expression of VEGF mRNA and VEGF in SKOV 3 cells The growth of xenografted tumor in nude mice was inhibited by reduced angiogenesis This provides an experimental basis for curing human ovarian cancer with antiangiogenesis method

关 键 词：实验研究 内皮生长因子 淋巴因子 催化RNA 卵巢肿瘤 实验性肿瘤 

分 类 号：R737.31[医药卫生—肿瘤]