基因沉默整合素连接激酶对滋养细胞生物学功能的影响

The suppressive effect of si ILK on proliferation, migration and invasion of human extravillous trophoblastic cells

目的:研究基因沉默整合素连接激酶(ILK)对人滋养细胞TEV-1细胞增殖、迁移和侵袭等的影响。方法:构建ILK-si RNA慢病毒载体后,转染人TEV-1细胞,分ILK基因沉默组(KD组)和空白质粒转染组(NC组)进行;q RT-PCR和Western blot检测m RNA和蛋白表达,MTT法、Transwell法和划痕实验等检测细胞的增殖、迁移和侵袭能力。结果:基因沉默ILK后,TEV-1细胞的ILK基因和蛋白的表达水平显著降低(P<0.05),证明干扰ILK表达成功。沉默ILK后,TEV-1细胞的增殖能力、侵袭能力、迁移能力均显著下降(P<0.05或 P<0.01)。ILK-si RNA转染TEV-1细胞后,MMP-9和p-GSK3β m RNA和蛋白水平显著下降(P<0.01)。结论:沉默ILK通过下调GSK3β和MMP-9的表达抑制人滋养细胞的增殖、迁移和侵袭。

Objective:To investigate the effects of integrin linked kinase(ILK)on the proliferation,migration and invasion of human extravillous trophoblastic cells(TEV-1).Methods:ILK-interferenced RNA was constructed and transfected into human TEV-1 cells,which was divided into experimental group(KD)and negative control group(NC);qRT-PCR and Western blot were used to detect the expression of ILK mRNA and protein.MTT assay,Transwell assay and Scarification test were used to detect the cell proliferation,migration and invasion.Results:Silenced ILK gene significantly reduced the ILK mRNA and protein expression levels of the TEV-1 cell(P<0.05),indicating that the transfection was successful.Silenced ILK gene significantly inhibited the proliferation,invasion(P<0.01)and migration of TEV-1 cells(P<0.05).Silenced ILK gene significantly decreased the mRNA and protein levels of MMP-9 and p-GSK3βin TEV-1 cells(P<0.01).Conclusion:These findings indicate that silenced ILK gene perturbs proliferation,migration and invasion may through down-regulated GSK3βand MMP-9 in TEV-1 cells.