CD47对正常妊娠不同阶段绒毛STOX1表达的影响

Effect of CD47 on STOX1 Expression in Villi at Different Stages of Normal Pregnancy

目的:通过比较研究CD47干预不同阶段正常妊娠绒毛前后STOX1的分子表达差异,探讨两者在滋养细胞的增殖、分化和凋亡过程中的相互关系和影响。方法:将试验对象早孕1组(妊娠5~7+6周,30例),早孕2组(妊娠8~11周,30例)绒毛进行细胞培养、染色体鉴别,随后将携带CD47基因的慢病毒同时转染干预早孕1组和2组。采用实时荧光定量PCR技术(RT-qPCR)检测各组绒毛在CD47干预前后STOX1基因的表达差异。各组细胞凋亡情况采用流式细胞仪检测分析。结果:各组细胞染色体核型检查提示均为正常健康人类染色体,未见明显数目和结构异常。绒毛细胞凋亡率随着孕周增加逐渐降低,且CD47转染的绒毛细胞凋亡率与未转染时比较明显降低,均有统计学意义。未干预前STOX1 mRNA表达在早孕1组相对较低,随着孕周进展表达相应增高。CD47干预后各组STOX1 mRNA表达仍呈现为随着孕周的增加而上升,且相对干预前各对应组明显升高(P<0.05)。结论:STOX1的分子表达水平伴随孕周的增加有逐渐升高趋势。CD47干预后表达这种趋势就更加明显,提示STOX1是CD47的下游调控因子,可能CD47通过正向干预STOX1的表达,从而参与对绒毛滋养细胞的增殖、分化和凋亡,影响绒毛的发生、发展进程,这对流产机制和保胎新方案的探索上具有重要意义。

Objective:To investigate the relationship and influence of CD47 on the proliferation, differentiation and apoptosis of trophoblasts by comparing the molecular expression difference of STOX1 before and after CD47 interferes with normal villi in different stages of pregnancy.Methods:Cell culture and chromosome identification were carried out on villi of test subjects in group 1 of early pregnancy(5~7+6 weeks, 30 cases) and group 2 of early pregnancy(8~11 weeks, 30 cases). Then lentivirus carrying CD47 gene was transfected to interfere with group 1 and group 2 of early pregnancy.Real-time fluorescence quantitative PCR(RT-qPCR) was used to detect the difference of STOX1 gene expression in villi before and after CD47 intervention.Apoptosis in each group was detected and analyzed by flow cytometry.Results:Karyotype examination of cells in each group showed that all cells were normal and healthy human chromosomes, and no obvious number and structural abnormalities were found. The apoptosis rate of villous cells decreased gradually with the increase of gestational age, and the apoptosis rate of villous cells transfected with CD47 was significantly lower than that of untransfected, with statistical significance. Before intervention, the expression of STOX1 mRNA was relatively low in group 1 of early pregnancy, and increased with gestational weeks.After CD47 intervention, the expression of STOX1 mRNA in each group still showed an increase with the increase of gestational weeks, and increased significantly compared with the corresponding groups before the intervention(P<0.05).Conclusion:The molecular expression level of STOX1 increases gradually with the increase of gestational age. The expression trend is even more obvious after CD47 intervention, suggesting that STOX1 is the downstream regulatory factor of CD47. It is possible that CD47 participates in the proliferation, differentiation and apoptosis of villous trophoblasts and affects the occurrence and development of villus by positively interfering with the expression of STOX1.This is of great significance to the exploration of the abortion mechanism and the new scheme of fetus protection.