微电场通过非整合素途径活化粘着斑激酶促进滋养细胞迁移/侵袭研究

Micro-electrical field may activate FAK through non-integrin pathway to promote trophoblast cells migration/invasion

目的探讨生理性微电场促进体外培养的人胎盘滋养细胞迁移/侵袭功能是否与滋养细胞表面整合素(integrin)表达有关。方法用150 m V/mm的直流微电场刺激滋养细胞,时间分别为5、10和15 h,测定其迁移情况并观察细胞形态变化。Western blot检测刺激前后1 h内粘着斑激酶FAK活化情况和细胞表面integrinα1、integrinα5、integrinαV和integrinα1蛋白表达水平。结果在含有10%胎牛血清的培养基中,150 m V/mm电场刺激下滋养细胞向负极定向迁移,迁移速度和距离较对照组明显增加(P=0.021),胞体拉长,垂直于电场方向排列;胞内FAKTyr397位点于刺激后5、10、30、60 min内迅速活化并逐渐加强(P<0.05);刺激前后滋养细胞表面integrinα1、integrinα5、integrinαV、和integrinα1蛋白表达水平无明显改变(P>0.05)。结论生理性直流微电场可能通过非整合素途径活化FAK从而促进滋养细胞迁移/侵袭功能,但其详细机制仍需进一步研究。

Objective To investigate whether the effect of physiological micro-electrical field on migration/invasion of trophoblast cells in vitro was associated with integrins expression of the ceils. Methods The trophoblast cells were exposed to the direct cur- rent electrical field （DC-EF） at 150 mV/mm for 5,10 and 15 hours. Cell images and migration distance were recorded with continuous photographing and analyzed by image analyzer. The activation of FAK at 5,10,30 and 60 min of DC-EF stimulation were measured and the expression levels of integrinα1 ,integrinα5, integrinαV and integrinα1 were detected by using Western blot assay. Results The application of 150 mV/mm DC electrical stimulation ,trophoblast cells cultured in media containing 10% calf serum showed a cathode migration. The migration velocity and distance were obviously increased when compared to the control group without EF stimulation （ P = 0. 021 ）. The cells exposed to the EF also showed elongation and perpendicular orientation, while control ceils that were not subjected to EF showed no such responsiveness. Compared with the non-EF stimulation controls, trophoblasts under EF stimulation had a quickly activation of FAKTyr397 location within 60 min （ P 〈 0. 05 ）, while no significant changes in expressions of integrinα1, integrinα5, integrinα V and integrinα1 were observed （ P 〉 0.05 ）. Conclusion The physiological direct current electrical field may activate FAK by non-integrin pathway to promote trophoblast cell migration/invasion function. However, its detailed mechanism needs further studies.