CAJ | 학술논문

目的：探讨滋养细胞迁移/侵袭相关促分裂原活化蛋白激酶/细胞外信号调节蛋白激酶(MAPK/ERK),磷酸肌醇-3激酶/丝氨酸苏氨酸蛋白激酶(PI3K/Akt)和蛋白酪氨酸激酶/信号转导和转录活化因子3(JAK/STAT3)信号传导通路在微电场刺激下的活化水平。方法将胎盘滋养细胞用场强为150 mV/mm 的直流微电场加以刺激,通过蛋白质印迹法(Western blot)测定电刺激前和后各时间点滋养细胞 MAPK/ERK、PI3K/Akt 和 JAK/STAT3信号传导通路相关活性分子的活化水平。滋养细胞在加电前分别用 MAPK 抑制剂 PD980599(100μmol/L)和 Akt 抑制剂 LY294002(20μmol/L)处理1 h,在上述相应含抑制剂的培养基中用场强为150 mV/mm 直流微电场刺激5 h,观察抑制剂对微电场刺激细胞行为的影响。结果Western blot 结果显示,滋养细胞在150 mV/mm 微电场作用下,p42/44 MAPK Thr202/Thr204位点、Akt Ser473位点于5 min 开始活化,10~60 min 内逐渐加强,STAT3 S727位点在微电场作用下上述各时间点无明显磷酸化水平改变。相应信号通路抑制剂处理后滋养细胞对微电场刺激的应答反应均受到明显抑制。结论微电场对滋养细胞迁移/侵袭功能的影响与 MAPK/ERK、PI3K/Akt 信号通路的活化有关。
목적：탐토자양세포천이/침습상관촉분렬원활화단백격매/세포외신호조절단백격매(MAPK/ERK),린산기순-3격매/사안산소안산단백격매(PI3K/Akt)화단백락안산격매/신호전도화전록활화인자3(JAK/STAT3)신호전도통로재미전장자격하적활화수평。방법장태반자양세포용장강위150 mV/mm 적직류미전장가이자격,통과단백질인적법(Western blot)측정전자격전화후각시간점자양세포 MAPK/ERK、PI3K/Akt 화 JAK/STAT3신호전도통로상관활성분자적활화수평。자양세포재가전전분별용 MAPK 억제제 PD980599(100μmol/L)화 Akt 억제제 LY294002(20μmol/L)처리1 h,재상술상응함억제제적배양기중용장강위150 mV/mm 직류미전장자격5 h,관찰억제제대미전장자격세포행위적영향。결과Western blot 결과현시,자양세포재150 mV/mm 미전장작용하,p42/44 MAPK Thr202/Thr204위점、Akt Ser473위점우5 min 개시활화,10~60 min 내축점가강,STAT3 S727위점재미전장작용하상술각시간점무명현린산화수평개변。상응신호통로억제제처리후자양세포대미전장자격적응답반응균수도명현억제。결론미전장대자양세포천이/침습공능적영향여 MAPK/ERK、PI3K/Akt 신호통로적활화유관。
Objective To investigate the effect of small direct-current electrical stimulation modulating activaty of p42/44 MAPK,PI3K/Akt and JAK/STAT3 signaling pathways relating to migration/invasion of trophoblast cells.Methods The tropho-blast cells (HTR-8/SVneo)were exposed to the DC-EF at 1 50 mV/mm for 0,5,10,30 and 60 min or 5,10 and 1 5 h.Phosphoryla-tion or expression of MAPK/ERK,PI3K/Akt,JAK/STAT3 were measured by Western blot analysis.MAPK and Akt inhibitors were used for treating the trophoblasts and then the effect of EF stimulation on the cell behaviors were observed.Results EF-stim-ulated trophoblast cells demonstrated p42/44 MAPK (Thr202/ Thr204)and Akt Ser473 sites were quickly phosphorylated within 5 min and then became intensively gradually within 60 min.There was no significant change of STAT3 S727 phosphorylation at the above time points for the EF-stimulated trophoblast cells.Conclusion The activation of p42/44 MAPK and PI3K/Akt signaling pathways might be involved in the DC-EF mediated directed migration of trophoblasts.