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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 effects of connective tissue growth factor (CTGF) on the expression of matrix metalloproteinase-9 (MMP-9), vascular endothelial growth factor (VEGF) and cell proliferation in human pancreatic cancer cell line SW1990 Mechanism of CTGF in Invasion and Metastasis of Pancreatic Cancer. Methods: The recombinant plasmid containing CTGF was transfected into human pancreatic cancer cell line SW1990 by lipofectamine. The positive cells were screened by G418 and identified by RT-PCR and Western blotting. The positive cells were detected by RT-PCR and Western blotting Cloning of MMP-9 and VEGF expression changes; thiazolyl tetrazolium (MTT) colorimetric detection of positive cell clone proliferation activity. Results: Positive SW1990 cell clones stably expressing high level of CTGF were successfully established. Compared with the control group, the expression of MMP-9, VEGF mRNA and protein in positive clones was significantly increased and the cell proliferation activity was also significantly increased. CONCLUSION: CTGF transfection can significantly increase the expression of MMP-9 and VEGF and promote the proliferation of pancreatic cancer cells. CTGF may be a potential target for gene therapy of pancreatic cancer.