CTGF基因转染对胰腺癌细胞MMP-9、VEGF表达及增殖的影响

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。

Effect of gene CTGF transfection on the expression of MMP-9 and VEGF and proliferation in human pancreatic cancer cells

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.