pegfp-n1-tgfβ1重组质粒对结肠癌细胞增殖、细胞周期及侵袭性的 .doc

pEGFP-N1-TGF-β1重组质粒对结肠癌细胞增殖﹑细胞周期及侵袭性的影响 于 宏姜 政王丕龙【摘要】 目的 探讨转化生长因子β1（TGF-β1）的真核表达载体对结肠癌细胞增殖﹑细胞周期及侵袭性的影响。方法 将构建的重组质粒pEGFP-N1-TGF-β1转染结肠癌细胞SW480中，应用荧光显微镜RT-PCR和Western blot检测TGF-β1基因的表达情况MTT法检测细胞生长情况；流式细胞仪分析SW480细胞周期变化；Transwell侵袭试验验证TGF-β1对细胞侵袭能力的影响。结果 转染重组质粒pEGFP-N1-TGF-β1后，通过荧光显微镜观察到绿色荧光的表达，RT-PCR和 Western blot检测到TGF-β1基因的表达增强细胞周期分析显示TGF-β1使细胞停留在G1期，S期细胞明显减少SW480细胞的增殖受到明显抑制Transwell侵袭试验SW480细胞的侵袭能力明显增强。结论 TGF-β1能抑制SW480细胞的增殖，使细胞停留在G1期，同时能增强其侵袭能力。免疫抑制/逃逸、增加血管生成增加肿瘤细胞与胞外基质的相互作用。 【关键词】转化生长因子-β1；SW480细胞；结肠癌；侵袭 【中图分类号】　　　　　 【文献标码】　 The effect of transforming growth factor-β1 on cell cycleroliferation and invasive capacity of human colon cancer cells YU hong, Jiang zheng， Wang Pilong (Department of Gastroenterology, The First Affiliated Hospital, Chongqing Medical University,Chongqing, 400016, China) 【Abstract】Objective investigate the effect of transforming growth factor-β1 (TGF-β1) on cell cycle, cell proliferation, and invasive capacity of human colon cancer cells of the line SW480, in order to lay the foundation for elucidating effects and its machaniam of TGF-β1 in colon carcinogenesis and development, as well as constructing the correspondence genetherapy with RNA interference. Methods: The recombinant expressing plasmids (pEGFP-N1-TGF-β1) was constructed and transfected into human colon cancer cells of the line SW480 cells by Lipofeetamine 2000. The expression of TGF-β1mRNA and protein in SW480 cells were detected by fluorescent microscopy, RT-PCR and Westen blot respectively. The cell proliferation of SW480 cells was tested by MTT assay, the cell cycle was analyzed by flow cytometry, and the invasion ability of SW480 cells were investigated by transwell-matrigel invasion chambers. Results: After transfected into SW480 cells, both mRNA and protein levels of TGF-β1 were effectively increased. Compared with the control group, the ability of cell proliferation transfected with pEGFP-N1-TGF-β1 was decreased significantly (P0.05), the inhibition rates were increas