微电场通过活化黏着斑激酶信号通路促进缺氧条件下绒毛外滋养细胞的迁移/侵袭  

Small direct-current electrical stimulation could improve migration / invasion of trophoblast cell under hypoxic condition by activating FAK signal pathway

作　　者：张娟[1] 李明勇[1] 贺元[1] 黄文芳[1] 白怀[2] 范平[2] 

机构地区：[1]四川省医学科学院.四川省人民医院检验科,成都610072 [2]四川大学华西第二医院.西部妇幼医学研究院,成都610041

出　　处：《临床检验杂志》2015年第11期813-817,共5页Chinese Journal of Clinical Laboratory Science

基　　金：国家自然科学基金(30872774/H0418);四川省卫生和计划生育委员会基金(140071)

摘　　要：目的探讨微电场刺激及缺氧条件对黏着斑激酶(FAK)信号通路的作用及对人胎盘绒毛外滋养细胞系HTR-8/SVneo迁移/侵袭的影响。方法划痕实验分析绒毛外滋养细胞在缺氧条件下的迁移情况;western blot检测微电场刺激后绒毛外滋养细胞FAK Tyr397和p Y576/577位点及其下游柱蛋白(paxillin)分子磷酸化水平,并检测微电场刺激5、10 h后HTR-8/SVneo细胞基质金属蛋白酶-2(MMP-2)的表达水平;2、5、10μmol/L FAK抑制剂PF-573228处理HTR-8/SVneo细胞后观察细胞行为,并用western blot检测其MMP-2的表达水平。结果微电场刺激能改善缺氧条件下HTR-8/SVneo细胞的迁移速度;与对照组比较,缺氧条件下微电场刺激可促使FAK Tyr397位点磷酸化水平及paxillin分子表达水平明显升高(P均<0.05),而p Y576/577位点则无明显改变;经微电场刺激5、10 h后,HTR-8/SVneo细胞MMP-2表达水平均升高(P均<0.05);经FAK抑制剂处理后,HTR-8/SVneo细胞对微电场的应答反应明显受抑制,其MMP-2表达水平降低。结论微电场可能通过活化FAK信号通路促进缺氧培养绒毛外滋养细胞的迁移/侵袭。Objective To investigate whether small direct-current electrical stimulation could improve migration and invasion of tropho- blast cells （HTR-8/SVneo） under hypoxic condition by mediating focal adhesion kinase （FAK） signal pathway activation. Methods Trophoblast cell migration under hypoxic condition was analyzed by wound healing assay. The phosphorylations of FAK at Tyr397, p- FAK, pY576/577 and paxillin, their downstream protein, in trophoblast cells were assessed by western blot. Matrix metalloproteinase- 2 （MMP-2） expression in trophoblast cells were analyzed in 5 and 10 hours after electrical stimulation. The cell behavior and MMP-2 expression of trophoblast cells were examined after treatment of FAK inhibitor PF-573228 at the concentration of 2, 5 and 10 μmol/L, respectively. Results Electrical stimulation could improve trophoblast cell migration and invasion under hypoxic condition. Compared with non-stimulated controls, phosphorylations of FAK at Tyr397 and the downstream paxillin in the electric stimulated trophoblast cells under hypoxic condition significantly increased （P 〈 O. 05）. However, no significant change of FAK phosphorylation at pY576/577 site was observed. MMP-2 expression in the 5- and 10-hour electrical stimulated cells was significantly increased （P 〈 0.05 ）. The im- proved cell migration in response to electrical stimulation under hypoxic condition was inhibited after FAK inhibitor PF-573228 treat- ment and MMP-2 expression was decreased with increasing concentration of FAK inhibitor. Conclusion The small direct electrical stimulation could improve trophoblast cell migration and invasion under hypoxic condition via activating FAK signaling pathway.

关 键 词：微电场 黏着斑激酶 滋养细胞 缺氧 迁移 侵袭 

分 类 号：R714.21[医药卫生—妇产科学]