刘志勇,欧阳忠,邹小明
LIU Zhiyong1,OUYANG Zhong1,ZOU Xiaoming2(1. Department of Breast Surgery

• 1:赣南医学院第一附属医院乳腺外科
• 2:哈尔滨医科大学附属第二医院胃肠外科

摘要(Abstract)：

目的:探讨类表皮生长因子域7(EGFL7)基因沉默对胃癌裸鼠模型瘤内血管生成的影响。方法:构建EGFL7短发夹状RNA表达质粒并转染SGC-7901细胞(pshEGFL7组),以空质粒转染为对照组,观察两组皮下种植瘤生长曲线及体积;免疫组织化学法检测抗-CD34、血管内皮生长因子(VEGF)、血小板反应蛋白(TSP1)表达,RT-PCR检测MMP-2和TIMP2表达。结果:pshEGFL7组种植瘤体积为(1.86±0.65)cm3,微血管密度(MVD)为20.84±6.38,分级为1～2级;对照组种植瘤体积为(4.86±1.15)cm3,MVD为39.48±9.01,分级为3～4级;两组种植瘤体积、MVD和分级的差异有统计学意义,P值均<0.05;EGFL7组TSP1蛋白阳性表达,而VEGF蛋白为弱阳性或阴性表达,MMP-2mRNA表达下调,而TIMP2mRNA表达上调,与对照组比较差异有统计学意义,P值均<0.01。结论:EGFL7基因沉默可降低胃癌种植瘤血管生成,与调节MMP-2/TIMP2表达,影响VEGF/TSP1平衡有关。
Objective:To investigate the effect of epidermal growth factor-like domain 7 (EGFL7) gene silencing on angiogenesis of stomach carcinoma in nude mice and its mechanism. Methods:A short hairpin RNA (shRNA) vector targeting at EGFL7 gene was constructed and transfected into SGC-7901 cells (pshEGFL7 group),and cells transfected with blank plasmids served as control group. Positive clones were selected and transplanted in nude mice to produce tumor animal model. Growth curves and volumes of tumors were observed. After 8 weeks,EGFL7 mRNA and protein in transplanted tumor tissues were detected and graded,and CD34,VEGF and TSP1 tested with immunochemistry method,and MMP-2 and TIMP2 mRNA detected with RT-PCR. Results:In pshEGFL7 group,the tumor volume was (1.86±0.65)cm3,and microvessel density (MVD) was 20.84±6.38,while in control group,tumor volume was (4.86±1.15)cm3,and MVD was 39.48±9.01. The differences between the two groups were significant (P<0.05).TSP1 protein expressed positively,and VEGF protein presented weakly or negatively in pshEGFL7 group. The expression of MMP-2 mRNA decreased,while TIMP2 mRNA increased in pshEGFL7 group,and the differences were significant statistically compared with those of control group (P<0.01). Conclusions:Silencing of EGFL7 gene can dicrease angiogenesis of stomach cancer in nude mice,which might relate to regulation of expression of MMP-2/TIMP2 and influence on balancing of TSP1/VEGF.

关键词(KeyWords)： 基因沉默;血管;癌;胃;表皮生长因子;小鼠,裸
gene silencing; carcinoma; stomach; blood vessels; epidermal growth factor; mice,nude

Abstract:

Keywords:

基金项目(Foundation):

作者(Author): 刘志勇,欧阳忠,邹小明
LIU Zhiyong1,OUYANG Zhong1,ZOU Xiaoming2(1. Department of Breast Surgery

DOI: 10.19367/j.cnki.1000-2707.2010.03.015

参考文献(References)：

• [1]李春杰,魏品康.胃癌细胞增殖相关基因研究进展[J].世界华人消化杂志,2007(9):975-979.
• [2]Parker LH,Schmidt M,Jin SW,et a1.The endothelial-cell-derived secreted factor Egfl7regulates vascular tube formation[J].Nature,2004(6984):754-758.
• [3]Yu JY,DeRuiter SL,Turner DL.RNA interference by ex-pression of Short-interfering RNAs and hairpin RNAs in mammalian cells[J].Proc Nad Acad Sci USA,2002(99):6047-52.
• [4]Picuric S,Friedrich M,Oess S.Expression and purification of recombinant human EGFL7protein[J].Protein Expr Purif,2009(1):1-6.
• [5]Schmidt MH,Bicker F,Nikolic I,et a1.Epidermal growth factor-like domain7(EGFL7)modulates Notch signalling and affects neural stem cell renewal[J].Nat Cell Biol,2009(7):873-880.
• [6]李功杰,杨立,史晓林,等.原发性肝癌肿瘤新生血管形态特征与其DSA血供分型关系的研究[J].介入放射学杂志,2005(2):135-138.
• [7]K sem M,Tuncer I,Kotan C,et a1.Significance of VEGF and microvascular density in gastric carcinoma[J].Hepa-toastroenterology,2009(93):1236-1240.
• [8]郭炜,单保恩,董稚明,等.贲门腺癌中TSP1启动子区高甲基化与TGF-β1表达的相关性分析[J].肿瘤,2009(10):971-975.
• [9]Mroczko B,Groblewska M,Lukaszewicz-Zajac M.Pre-treatment serum and plasma levels of matrix metalloprotei-nase9(MMP-9)and tissue inhibitor of matrix metallopro-teinases1(TIMP-1)in gastric cancer patients[J].Clin Chem Lab Med,2009(9):1133-1139.

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