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作 者:倪效[1] 燕敏[1] 严超[1] 刘炳亚[1] 陈雪华[1] 瞿颖[1] 计俊[1] 俞焙秦[1] 朱正纲[1]
机构地区:[1]上海交通大学医学院附属瑞金医院普外科上海消化外科研究所,上海200025
出 处:《诊断学理论与实践》2008年第2期198-201,共4页Journal of Diagnostics Concepts & Practice
基 金:上海市科委基金项目(044119664)
摘 要:目的:研究血管内皮生长因子C(vascular endothelial growth factor-C,VEGF-C)与胃癌淋巴管生长间的关系。方法:采用逆转录-聚合酶链反应(RT-PCR),从人胃癌旁组织中扩增人VEGF-CcDNA全长,经T-A克隆酶切,鉴定并测序得到的完整序列后,进行亚克隆并构建pEGFP/VEGF-C真核表达载体,将其转染低表达VEGF-C的胃癌细胞株MKN45;另转染pEGFP空载体,作为对照组。通过体外G418抗生素筛选,得到高表达VEGF-C的肿瘤细胞克隆,用这种高表达VEGF-C的胃癌细胞与对照组细胞接种于裸鼠皮下(2组各为10只),观察肿瘤组织内淋巴管密度及裸鼠瘤体质量。结果:对照组和转染pEGFP/VEGF-C组肿瘤质量分别为(2.64±0.92)g、(5.55±0.81)g,差异有统计学意义(P<0.001);2组肿瘤组织内的淋巴管密度分别为9.77±1.12,13.87±1.29,差异有统计学意义(P<0.05)。结论:VEGF-C能在胃肿瘤中促淋巴管生长。Objective To investigate the correlation between VEGF-C and lymphatic vessel metastasis in gastric carcinoma. Methods Full length VEGF-C eDNA isolated from para-cancer gastric tissue was amplified by RT-PCR. The complete sequence obtained after T-A cloning, enzyme digestion identification and sequencing was subcloned and a pEGFP/ VEGF-C vector was constructed. Eukaryotic expression vector pEGFP/VEGF-C was transfected into human gastric carcinoma cell line MKN45 with low expression VEGF-C which served as study group, and empty vector pEGFP was transfected into MKN45 which served as a control group. Stable VEGF-C high-expression MKN45 clones were selected in vitro with G418, which were then combined and subcutaneously injected into nude mice to quantify the density of lymphatic vessel compared with MKN45 cells transfected only with empty vector. Results The weight of tumor in pEGFP/ VEGF-C tranfected group [(5.55±0.81) g] was significantly higher than that in the control group [(2.64±0.92) g] (P〈0.001). LVD (lymphatic vessel density) in pEGFP/VEGF-C transfected group (13.87±1.29) was also significantly higher than that in the control group (9.77±1.12) (P〈0.05). Conclusions In gastric carcinoma, VEGF-C may promote the growth of tumor through promoting the growth of lymphatic vessels.
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