摘要
目的探讨在体外转化生长因子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转染入结肠癌细胞系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侵袭实验验证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Westernblot检测到TGF-β1基因的转录及蛋白表达增强(P<0.05);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(P<0.05),且SW480细胞的增殖受到明显抑制;Transwell侵袭实验显示:SW480细胞的侵袭能力明显增强(P<0.01)。结论TGF-β1能抑制SW480细胞的增殖,使细胞停留在G1期,但同时能增强其侵袭能力。其作用可能与免疫抑制/逃逸、增加血管生成以及增加肿瘤细胞与细胞外基质的相互作用有关。
Objective To 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,for its role in colon carcinogenesis and development,as well as its application in gene therapy for colon cancer with RNA interference.Methods The recombinant expressing plasmids pEGFP-N1-TGF-β1 was constructed and transfected into SW480 cells by Lipofectamine 2000.The expression of TGF-β1 mRNA and protein in the transfected SW480 cells were detected by fluorescent microscopy,RT-PCR and Western blot analysis 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 the mRNA and protein levels of TGF-β1 were effectively increased （P0.05）.Compared with the control group,the ability of cell proliferation transfected with pEGFP-N1-TGF-β1 was decreased significantly （P0.05）,the inhibitory rates were increased from 60% to 80%,the cells cycle were arrested at G0/G1 stage,and the invasive ability were evidently increased （P0.01）.Conclusion TGF-β1 inhibits the cell proliferation and arrests cell cycle,but increases cell invasion ability in vitro.Its effects may be related to immunosuppression/escape,increased angiogenesis and reinforcement interaction of tumor cells with extracellular matrix.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2010年第9期943-946,共4页
Journal of Third Military Medical University
