mir-184在反复自然流产中的作用及机制研究

Accumlating evidence already shows that microRNAs are small non-coding RNA which are involved in cell differentiation,development and the occurrence and pathogenesis of many diseases. At present study, first, we did the micro-array for microRNA expression in deciduas and villi from recurrent spontanous abort patients.it showed that the expression profiles of microRNAs were significantly difference between that in deciduas or villi in normal abortion patients and that in deciduas or villi in recurrent spontanous abortion patients.Espacially, the expression of mir-184 both in deciduases and villi in recurrent spontanous abortion patients were significantly higher than that in deciduases and villi in normal abortion patients. Prediction of the target genes of mir-184 suggest that mir-184 was closely associated with the cell development, differentiation and immune response.In order to extensively to explore the function of mir-184 in recurrent spontanous abortion, further experiments including mir-184 function on proliferation, invision, apoptosis and protein secreting profile of trophoblast are going to be conducted.

microRNA是一类非编码的小分子RNA，广泛参与了细胞的分化、发育及各种临床疾病的发生发展。本课题我们首先对反复自然流产患者和正常妊娠妇女的绒毛和蜕膜组织进行microRNA 芯片分析，通过差异microRNA筛选及功能预测,我们发现mir-184在反复自然流产患者的绒毛和蜕膜组织中均显著高表达，且其靶基因分析显示mir-184与细胞发育、分化、免疫应答等功能密切相关。因此我们推测，mir-184在反复自然流产中可能起到关键调控作用。本研究拟在已有预实验的基础上，通过细胞生物学和分子生物学实验，进一步探讨mir-184对胎盘滋养细胞生物学行为的影响；并阐明mir-184引起的这些生物学功能改变的机制，即mir-184通过调控那些基因的表达来引起滋养细胞生物学行为变化。通过对以上问题的解决，进一步阐明反复自然流产的发病机制，为临床反复自然流产的治疗提供新的思路。

microRNA是一类非编码的小分子RNA，广泛参与了细胞的分化、发育及各种临床疾病的发生发展。本课题我们首先对原因不明反复自然流产患者和正常人流患者的绒毛和蜕膜组织进行microRNA表达芯片分析，发现在反复自然流产患者绒毛组织中上升的microRNA有3个下降的有3个；在蜕膜组织中我们检测到5个microRNA表达上升。之后，我们采用qRT-PCR方法扩大样本量对11个差异表达microRNA进行验证。综合芯片和qRT-PCR结果，我们发现hsa-miR-184同时高表达于反复自然流产患者的绒毛和蜕膜组织。本项目研究中，我们在已取得数据的基础上利用慢病毒载体构建了稳定高表达has-miR-184和整合空载的HTR8细胞系。表型分析发现，在HTR8细胞系中高表达hsa-miR-184可以显著抑制细胞增殖且可以促进凋亡。生物信息学分析预测结果显示，hsa-miR-184可能通过靶向ARHGDIA、SPHK2和SELS等基因促进细胞的凋亡；通过靶向GNB1、PDXK和LMO1来影响增殖。此外，我们对生物信息学预测的has-miR-184的靶基因之一AGO2进行了验证，发现hsa-miR-184可以直接靶向AGO2的3’-UTR来抑制AGO2蛋白的表达。