CAJ | 학술논문

目的：探讨成蛋白样3型蛋白（FM NL3）基因转染对结直肠癌SW480细胞上皮-间质转化（EM T ）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FMNL3 mRNA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48 h后SW480细胞中FMNL3 mRNA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。
목적：탐토성단백양3형단백（FM NL3）기인전염대결직장암SW480세포상피-간질전화（EM T ）화체외침습능력적영향。방법장대록색형광단백적중조 FMNL3표체질립pEGFP-N1/FMNL3전염지SW480세포중（이미전염질립적SW480세포작정상대조，전염공재질립적SW480세포위음성대조），형광정량PCR（RT-qPCR）화Western blot법검측중조질립전염전、후세포FMNL3 mRNA화단백적표체변화；재형광현미경하관찰FMNL3전염전、후SW480세포적형태변화；채용RT-qPCR화Western blot검측전염전、후SW480세포EMT상피표지물E-cadherin화간질표지물Vimentin적표체변화；용Transwell소실실험검측FMNL3기인전염대SW480세포적체외침습능력적영향。결과 FMNL3전염48 h후SW480세포중FMNL3 mRNA화단백표체수평명현제고（P＜0．01）；FMNL3단백표체우세포질중，전염 FMNL3기인후SW480세포적형태발생료명현적개변，유전형적원형、다변형변성사형병신출다개세장돌기적간충질양，정현전형적EMT형태학개변；전염FMNL3기인후，SW480세포중EMT상피표지물E-cadherin표체하조，간질표지물Vimentin표체상조；전염FMNL3기인후SW480세포적체외침습능력현저증강。결론 FMNL3기인가능통과촉진결직장암세포EMT이촉진기침습。
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<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 .