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作 者:姜建国[1] 焦霞[2] 朱晓蔚[2] 戴桂红[2] 肖蔚[2] 刘欣韵 窦荣荣 周彤敏 于鸿[2]
机构地区:[1]泰州市人民医院普外科,江苏泰州225300 [2]泰州市人民医院病理科,江苏泰州225300
出 处:《江苏大学学报(医学版)》2013年第5期377-381,共5页Journal of Jiangsu University:Medicine Edition
基 金:江苏省"333工程"培养资金资助项目(200924);南通大学自然科学基金资助项目(10Z088)
摘 要:目的:通过研究结缔组织生长因子(connective tissue growth factor,CTGF)基因对人胰腺癌SW1990细胞基质金属蛋白酶-9(MMP-9)、血管内皮生长因子(VEGF)表达及细胞增殖的影响,深入探讨CTGF在胰腺癌侵袭和转移中的作用机制。方法:经脂质体介导将含有CTGF重组表达质粒转染人胰腺癌SW1990细胞株,用G418筛选阳性细胞克隆及RT-PCR、蛋白质印迹法鉴定;采用RT-PCR和蛋白质印迹法检测阳性细胞克隆MMP-9及VEGF表达的改变;噻唑盐(MTT)比色法检测阳性细胞克隆的增殖活性。结果:成功建立稳定高表达CTGF的阳性SW1990细胞克隆,与对照组相比,阳性克隆MMP-9、VEGF mRNA及蛋白表达显著增高,细胞增殖活性也显著增加。结论:CTGF转染可显著增加胰腺癌细胞MMP-9、VEGF的表达并促进其细胞增殖,CTGF可能是胰腺癌基因治疗的潜在靶点。Objective: To investigate the effect of connective tissue growth factor(CTGF) on the proliferation in human pancreatic cancer cell line SW1990,with a focus on the expression of matrix metalloproteinase-9(MMP-9) and vascular endothelial growth factor(VEGF).This study was designed to explore the underlying mechanism for the role of CTGF in the development of pancreatic cancer.Methods: pcDNA3.0CTGF and pcDNA3.0 were transfected into SW1990 cells through lipofectamine and positive clones were screened by G418.RT-PCR and Western blotting were employed to identify mRNA and protein expression of CTGF in SW1990 cells,respectively.The expression of MMP-9 and VEGF in positive clones was detected by RT-PCR and Western blotting.Cell viability was assessed by dimethylthiazol diphenyl tetrazoliumbromide(MTT) method.Results: Positive clones C-5 and C-28 with CTGF over-expression were successfully established.Compared with non-transfected control group,the expressions of MMP-9 and VEGF in C-5 and C-28 was increased significantly,and the proliferation level of C-5 and C-28 was increased significantly.Conclusions: CTGF transfection could effectively enhance the expression of MMP-9 and VEGF and the proliferation in SW1990 cells which suggested a potential role for CTGF in gene therapy of pancreatic cancer.
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