【摘要】 目的探讨成蛋白样3型蛋白(FMNL3)基因转染对结直肠癌SW480细胞上皮-间质转化(EMT)和体外侵袭能力的影响。方法将带绿色荧光蛋白的重组FMNL3表达质粒pEGFP-N1/FMNL3转染至SW480细胞中(以未转染质粒的SW480细胞作正常对照,转染空载质粒的SW480细胞为阴性对照),荧光定量PCR(RT-qPCR)和Western blot法检测重组质粒转染前、后细胞FMNL3mRNA和蛋白的表达变化;在荧光显微镜下观察FMNL3转染前、后SW480细胞的形态变化;采用RT-qPCR和Western blot检测转染前、后SW480细胞EMT上皮标志物E-cadherin和间质标志物Vimentin的表达变化;用Transwell小室实验检测FMNL3基因转染对SW480细胞的体外侵袭能力的影响。结果 FMNL3转染48h后SW480细胞中FMNL3mRNA和蛋白表达水平明显提高(P<0.01);FMNL3蛋白表达于细胞质中,转染FMNL3基因后SW480细胞的形态发生了明显的改变,由典型的圆形、多边形变成梭形并伸出多个细长突起的间充质样,呈现典型的EMT形态学改变;转染FMNL3基因后,SW480细胞中EMT上皮标志物E-cadherin表达下调,间质标志物Vimentin表达上调;转染FMNL3基因后SW480细胞的体外侵袭能力显著增强。结论 FMNL3基因可能通过促进结直肠癌细胞EMT而促进其侵袭。更多还原

【Abstract】 Objective To investigate the impact of FMNL3gene transfection on the epithelial-mesenchymal transition(EMT) and the in vitro invasive potential of colorectal carcinoma SW480cells.Methods The recombination FMNL3expression plasmid with EGFP was transfected into SW480cells(SW480cells without transfected plasmid as the normal control group and SW480cells with empty vector as the negative control group).The changes of FMNL3mRNA and protein expression were detected by real-time PCR and Western blot after transfection of 48hwith recombination FMNL3expression plasmid.The changes of SW480cell morphology were detected by the fluorescence microscope after FMNL3transfection.The changes of EMT epithelial and mesenchymal markers in SW480cells were detected by real-time PCR and Western blot after FMNL3transfection.The Transwell chamber invasion assay in vitro was used to investigate the effects of FMNL3gene trasfection on the invasive potential of SW480cells.ResultsThe levels of FMNL3mRNA and the protein expression were significantly increased at 48hafter transfection with recombination FMNL3expression plasmid(P<0.01);the FMNL3protein was expressed within cytoplasma,the morphology of SW480cells after FMNL3transfection showed significantly changes from round and polygon to spindle and stretched out the multiple long and thin prominences as mesenchyma,which appeared the typical EMT change;after FMNL3gene transfection,the expression of EMT epithelial marker E-cadherin was down-regulated and the expression of mesenchymal marker Vimentin was up-regulated in SW480 cells;the in vitro invasive potential of SW480cells was remarkably enhanced after FMNL3transfection.Conclusion FMNL3gene may promote its invasion by promoting EMT of colorectal carcinoma.更多还原