早期自然流产患者绒毛KiSS-1、MMP-9和MMP-2基因及其蛋白表达

目的研究早期自然流产患者绒毛肿瘤转移抑制基因KiSS-1与基质金属蛋白酶(MMP-9、MMP-2)mRNA及蛋白的表达,探讨其与自然流产的关系。方法采用半定量RT-PCR、免疫印迹法检测30例正常早期妊娠和30例早期自然流产患者绒毛KiSS-1、MMP-9和MMP-2的mRNA及蛋白水平,用明胶酶谱分析法检测上述样本孵育液MMP-9、MMP-2的酶活性。结果(1)自然流产患者绒毛组织中MMP-9及MMP-2mRNA表达水平低于正常早期妊娠组(P<0.05),KiSS-1mRNA的表达高于正常早期妊娠组差异有统计学意义(P<0.05);(2)Westernblotting分析显示自然流产患者绒毛MMP-9、MMP-2蛋白表达低于正常早期妊娠差异有统计学意义(P<0.05),KiSS-1蛋白表达高于正常早期妊娠差异有统计学意义(P<0.05);(3)明胶酶谱分析结果显示,自然流产患者绒毛MMP-9、MMP-2酶活性低于正常早期妊娠者差异有统计学意义(P<0.05)。结论自然流产患者绒毛促浸润基因MMP-9、MMP-2mRNA及其蛋白表达水平降低,同时抑制浸润基因KiSS-1mRNA及其蛋白表达增加,造成滋养细胞浸润能力下降,可能与自然流产的发生有关。

Expression of mRNA and protein of KiSS-1,MMP-9 and MMP-2 in villi of first trimester spontaneous abortion

Objective:To investigate the mRNA and protein expression of metastasis suppressor gene KiSS-1, matrix metalloproteinase-9(MMP-9) and MMP-2 in villi of first trimester spontaneous abortion.Methods:Expression of KiSS-1,MMP-9 and MMP-2 mRNA and protein in villi of first trimester spontaneous abortion (abortion group,n=30) and normal first trimester pregnant women(control group,n=30) was detected by Semi-quantitative RT-PCR ,Western blotting and gelatin zymography.Results:(1)Expression of MMP-9 and MMP-2 mRNA was significantly lower in villi of abortion group compared with that of control group(P<0.05).Expression of KiSS-1mRNA increased significantly in villi of abortion group(P<0.05);(2)Western blotting analysis demonstrated that expression of MMP-9 and MMP-2 protein in villi of abortion group was significantly lower than that of control group(P<0.05).Expression of KiSS-1 protein in villi of abortion group was significantly higher than normal(P<0.05);(3)Gelatin zymographic analysis showed enzyme activity of MMP-9 and MMP-2 in harvested villi culture medium of abortion group was significantly lower than that of control group(P<0.05).Conclusion:This study showed that MMP-9,MMP-2 and KiSS-1 may play an important role in the invasion of human placenta trophoblast.It may correlate with occurrence of first trimester spontaneous abortion.