FMNL3基因转染对结直肠癌SW480细胞EMT和体外侵袭能力的影响  

Influence of FMNL3 gene transfection on epithelial-mesenchymal transition and in vitro invasion potential of colorectal carcinoma SW480 cells

作　　者：曾元凤[1] 肖移生[2] 罗小江[3] 胡国柱[4] 路名芝[1] 吴晓牧[5] 辛洪波[6] 

机构地区：[1]江西省人民医院病理科,江西南昌330006 [2]江西中医药大学基础医学院形态学教研室,江西南昌330004 [3]江西省人民医院普外科,江西南昌330006 [4]江西省人民医院干细胞重点实验室,江西南昌330006 [5]江西省人民医院神经内科,江西南昌330006 [6]南昌大学转化医学研究院,江西南昌330031

出　　处：《重庆医学》2014年第12期1460-1462,1467,共4页Chongqing medicine

基　　金：国家自然科学基金资助项目(81201972;81260327)

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。Objective To investigate the impact of FMNL3 gene transfection on the epithelial-mesenchymal transition（EMT） and the in vitro invasive potential of colorectal carcinoma SW 480 cells .Methods The recombination FMNL3 expression plasmid with EGFP was transfected into SW480 cells（SW480 cells without transfected plasmid as the normal control group and SW480 cells with empty vector as the negative control group） .The changes of FMNL3 mRNA and protein expression were detected by real-time PCR and Western blot after transfection of 48h with recombination FMNL3 expression plasmid .The changes of SW480 cell mor-phology were detected by the fluorescence microscope after FMNL 3 transfection .The changes of EMT epithelial and mesenchymal markers in SW480 cells were detected by real-time PCR and Western blot after FMNL3 transfection .The Transwell chamber inva-sion assay in vitro was used to investigate the effects of FMNL 3 gene trasfection on the invasive potential of SW480 cells .Results The levels of FMNL3 mRNA and the protein expression were significantly increased at 48h after transfection with recombination FMNL3 expression plasmid（P＆lt;0 .01）;the FMNL3 protein was expressed within cytoplasma ,the morphology of SW480 cells after FMNL3 transfection 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 FMNL3 gene transfection ,the expression of EMT epi-thelial 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 SW480 cells was remarkably enhanced after FMNL3 transfection .Conclusion FMNL3 gene may promote its invasion by promoting EM T of colorectal carcinoma .

关 键 词：结直肠肿瘤 成蛋白样3型蛋白 上皮-间质转化 侵袭 

分 类 号：R735.34[医药卫生—肿瘤]