restin基因转染对裸鼠胃癌移植瘤生长的影响  

Effect of restin gene transfection on human gastric carcinoma xenograft growth in nude mice

作　　者：张建永[1] 李锦毅[2] 李红玉[1] 戴聪[1] 马春雷[1] 王向党[2] 

机构地区：[1]辽宁医学院,辽宁省锦州市121001 [2]中国人民解放军武警总医院,北京市100039

出　　处：《世界华人消化杂志》2008年第33期3713-3718,共6页World Chinese Journal of Digestology

基　　金：国家自然科学基金资助项目;No.30672771~~

摘　　要：目的:探讨restin基因转染对胃癌细胞BGC-803裸鼠移植瘤生长抑制作用及对肿瘤血管生成的影响.方法:将重组质粒pEGFP-restin转染BGC-803细胞,以绿色荧光蛋白示踪其对胃癌细胞BGC-803生长的影响;RT-PCR鉴定目的基因的表达;通过裸鼠移植瘤实验比较restin对裸鼠移植瘤的生长抑制作用,免疫组织化学染色方法观察微血管密度(MVD)和血管内皮生长因子(VEGF)的表达.结果:重组质粒经RT-PCR扩增后在600bp处稳定表达;将pEGFP-restin重组质粒转染胃癌细胞BGC-803,荧光显微镜下见转染组细胞发出绿色荧光,肿瘤细胞形态不一;裸鼠移植瘤实验显示,与对照组(空载体组和未转染组)比较,转染组胃癌细胞生长速度缓慢,肿瘤体积小(P<0.01);空载体组与转染组的抑瘤率分别为3.60%、29.02%;免疫组化显示,转染组肿瘤微血管密度减少(4.25±0.29vs9.79±0.94,10.34±1.22,均P<0.05),转染组VEGF表达降低(12.24±3.45vs44.52±9.70,39.76±6.38,均P<0.05).结论:restin基因转染可抑制裸鼠移植瘤生长,影响肿瘤新生血管形成可能是其抗肿瘤作用之一.AIM：To investigate the inhibitory effect of restin transfection on the growth of human gastric cancer BGC-803 tumor xenograft in nude mouse model with the localized expression of restin and to explore its effect on angiogenesis.METHODS：Gastric cancer BGC-803 cell line was transfected with recombinant pEGFP-restin through LipofectamineTM2000 mediation.The effect on BGC-803 growth was observed by enhanced green fluorescent proteins assay.The expression of restin gene was detected using re-striction endonuclease analysis and RT-PCR;cell cycle was observed using flow cytometry（FCM）.Through in vitro study,growth inhibition of the restin on the cancer was observed in nude mice.Microvessel density（MVD）and expression of vascular endothelial growth factor（VEGF）were assessed by immunohistochemistry.RESULTS：The results of identification by restriction endonuclease analysis and RT-PCR demonstrated that restin gene was stably expressed in BGC-803 cells transfected with pEGFP-restin and that significant expression of EGFP was observed in these cells with irregular-ity.Slower growth speed and smaller volume were found in pEGFP-restin thansfecting cell group than in control group（P〈0.01）.The inhibition rates of tumor in empty vector and pEGFP-restin thansfecting cell group were 3.60% and 29.02%,respectively.The MVD was lower in pEGFP-restin thansfecting cell group than in control group（4.25±0.29 vs 9.79±0.94,10.34± 1.22,both P〈0.05）;the expression of VEGF was down-regulated compared with control group（12.24±3.45 vs 44.52±9.70,39.76±6.38,both P〈 0.05）.CONCLUSION：Restin transfection could inhibit BGC-803 cell growth in nude mice,which may be related to inhibition of angiogenesis in gastric cancer.

关 键 词：restin基因 转染 胃肿瘤 微血管密度 血管内皮生长因子 

分 类 号：R735.2[医药卫生—肿瘤]