GPR30在妊娠期胎盘中的表达及其对滋养细胞侵袭能力影响的研究

The research about the expression of GPR30 protein in human placenta and its effect on human trophoblast cells invasion

目的:探讨G-蛋白偶联受体30(GPR30)在妊娠期胎盘中的表达,以及其对人胎盘滋养细胞侵袭力的影响。方法:免疫组化法检测早孕期绒毛、正常产妇胎盘和重度子痫前期患者胎盘组织中GPR30表达。用17-β-雌二醇(17β-E2)、GPR30激动剂G1和阻滞剂G15预处理体外培养绒毛组织和人绒毛外滋养细胞株HTR8/SVneo。显微镜下观察体外绒毛组织滋养细胞生长侵袭范围,Transwell侵袭实验检测HTR8/SVneo细胞侵袭能力。免疫荧光法检测绒毛组织和HTR8/SVneo细胞中GPR30蛋白表达。Western blot法检测HTR8/SVneo细胞中GPR30和MMP-9蛋白表达。结果:GPR30在早期绒毛组织和正常末期胎盘滋养细胞上都有表达,且早孕期的表达水平高于正常末期胎盘,但在重度子痫前期胎盘上GPR30蛋白表达明显减少。E2及GPR30激动剂G1可增加滋养细胞的侵袭能力,阻滞剂G15则可下调其侵袭性;E2、G1可诱导滋养细胞GPR30蛋白表达上调,而G15则下调其表达。GPR30蛋白水平与侵袭相关蛋白MMP-9表达水平有相关性。结论:GPR30可能参与人类滋养细胞侵袭力的调节,对滋养细胞的侵袭力有正性促进作用。

Objective：To characterise the expression of G-Protein Coupled Receptor 30 （GPR30） in human placenta, and to determine the role of GPR30 in trophoblast invasion. Meth- ods：Placentas from first trimester, normal term and pregnancies complicated with pre-eclampsia were used in the study. The effect of 17 ^-estradiol （ E2 ）, GPR30 agonist G1 and GPR30 inhibitor G15 on invasion of first trimester villous explants and human trophoblast cell line HTR8/SVneo were determined using in vitro invasion assays. Immunofluorescence and Western blot were used to investigate the GPR30 protein expression. Results：GPR30 protein expression was significantly lower in term placentas from pregnnacies complicated by pre-eclampsia compared to placentas from first trimester and normal term pregnancies. E2 and G1 treatment significantly increased the invasiveness of trophoblast cells. In contrast, the antagonist G15 significantly decreased tropho- blast invasion. In addition, E2 and G1 treatment increased while G15 decreased GPR30 protein ex- pression. Meanwhile, the results also implied that GPR30 involves in regulating the expression of MMP-9. Conclusion： GPR30 plays a role in trophoblast invasion, the up-regulation of GPR30 pro- tein level could increase trophoblast invasion.