CAJ | 학술논문

目的探讨成纤维细胞生长因子2(FGF2)逆转转化生长因子-β1(TGF-β1)诱导的肺癌上皮细胞间质转化(EMT)的机制.方法用细胞划痕和Transwell侵袭试验检测细胞运动和侵袭能力变化,用免疫荧光和Western blot法检测逆转过程中关键分子的表达变化.用特异性通路抑制剂分析FGF2逆转EMT发挥作用的的信号通路.结果 TGF-β1作用72 h后,肺癌上皮细胞发生EMT,细胞运动、侵袭能力增强(200.0±12.5,P＜0.01).联合FGF2继续培养48h之后,细胞各项指标恢复至对照水平,运动、侵袭能力下降(50.0±5.5,P＜0.01).利用特异性通路抑制剂LY294002和PD98059后结果显示,抑制丝裂原激活的蛋白激酶/细胞外调节蛋白激酶(MAPK/ERK)通路可以明显阻断FGF2对于TGF-β1诱导EMT的逆转作用,而激活MAPK/ERK通路可以降低TGF-β1对Smad2的磷酸化.结论 FGF2通过MAPK/ERK通路逆转TGF-β1诱导的肺癌上皮细胞EMT.
목적 탐토성섬유세포생장인자2(FGF2)역전전화생장인자-β1(TGF-β1)유도적폐암상피세포간질전화(EMT)적궤제.방법 용세포화흔화Transwell침습시험검측세포운동화침습능력변화,용면역형광화Western blot법검측역전과정중관건분자적표체변화.용특이성통로억제제분석FGF2역전EMT발휘작용적적신호통로.결과 TGF-β1작용72 h후,폐암상피세포발생EMT,세포운동、침습능력증강(200.0±12.5,P＜0.01).연합FGF2계속배양48h지후,세포각항지표회복지대조수평,운동、침습능력하강(50.0±5.5,P＜0.01).이용특이성통로억제제LY294002화PD98059후결과현시,억제사렬원격활적단백격매/세포외조절단백격매(MAPK/ERK)통로가이명현조단FGF2대우TGF-β1유도EMT적역전작용,이격활MAPK/ERK통로가이강저TGF-β1대Smad2적린산화.결론 FGF2통과MAPK/ERK통로역전TGF-β1유도적폐암상피세포EMT.
Objective To investigate the effects of fibroblast growth factor 2 (FGF2) reversing transforming growth factor β1 (TGF-β1)-induced epithelial-mesenchymal transition (EMT) of lung cancer cell A549.Methods Cell scratch and Transwell methods were used to examine the migration and invasion ability of cancer cells.The expression changes of key EMT and MET molecular maker E-cadhefin,Vimentin,N-cadherin,and Snail were measured by using immunofluorescence and Western blotting.Signaling pathways were analyzed by using selective inhibitors LY294002 and PD98059.Results The morphological changes and the epithelial and mesenchymal markers induced by TGF-β1 were reversed by FGF2 to control levels.FGF2 also decreased the migration and invasion ability of A459 cells (50.0 ± 5.5,P ＜ 0.01).Signaling pathways analyzed by selective inhibitors showed that the inhibition of MAPK/ERK kinase pathway could substantially block the reversal effects of FGF2,while the activation of MAPK/ERK kinase pathway resultd in Smad2 dephosphorylation.Conclusion These findings indicate that the TGF-β1-induced EMT is reversed by FGF2 through the MAPK/ERK kinase pathway.