CCL28通过增强MMP-2活力促进滋养细胞浸润功能

The chemokine CCL28 enhances human trophoblast cell invasion by promoting MMP-2 activity

目的:探究趋化因子CCL28对滋养细胞生物学行为的影响及其调控机制。方法:分离培养并鉴定原代人早孕蜕膜基质细胞(DSCs),ELISA法检测人DSCs与人正常绒毛膜滋养细胞系HTR8/Svneo的CCL28的分泌。采用噻唑兰(MTT)、Annexin V-FITC、Transwell方法检测人工合成的CCL28对HTR8/Svneo细胞的增殖、凋亡及浸润能力的影响。RT-PCR及明胶酶谱法阐明HTR8/Svneo细胞浸润功能改变的机制。结果:HTR8/Svneo细胞与蜕膜基质细胞均能分泌CCL28,两者共培养能促进CCL28的分泌(P<0.01)。人工合成的CCL28不影响滋养细胞的增殖(P>0.05)和凋亡(P>0.05),但能增强细胞的浸润能力(P<0.01),且主要通过受体CCR10介导。RT-PCR与明胶酶谱法提示,CCL28通过提高滋养细胞内MMP-2活力来增强其浸润能力。结论:CCL28通过与受体CCR10的结合,以自分泌或旁分泌的方式增强滋养细胞MMP-2活力来促进滋养细胞的浸润功能。

Objective：To investigate the effect of chemokine CCL28 on trophoblast cells invasion.Methods：Decidual stromal cells（DSCs） were isolated,and indentified from healthy women of early pregnancy.Protein expression of CCL28 was detected in DSCs,HTR8/Svneo cells and their co-culture supernatants by enzyme-linked immunosobent assay.The effect of rhCCL28 on HTR8/Svneo cells proliferation was assessed by the MTT assay.The flow cytometry was used to detect the effect of rhCCL28 on HTR8/Svneo cells apoptosis using annexinV-FITC/PI staining.The effect of rhCCL28 on HTR8/Svneo cells invasion was detected by Transwell chamber.MMP-2 and MMP-9 activity in the supernatants of HTR8/Svneo cells was treated by rhCCL28 in the invasion assay detected by RT-PCR and gelatin zymography.Results：The DSCs and HTR8/Svneo cells could secretion CLL28 inceased with time,and their co-culthure could promote the secreted of CCL28 greatly（P〈0.01）.RhCCL28 did not affect the proliferation（P＆gt;0.05） or apoptosis of HTR8/Svneo cells（P＆gt;0.05） but promoted its invasion（P〈0.01） in a proper concentration through binding with its CCR10 receptotor mainly.RhCCL28 promoted cell invasion by enhancing MMP-2 activity.Conclusion：CCL28 could enhance trophoblast cell invasion by binding with receptor CCR10 through promoting the activity of MMP-2 of trophoblast cell.