摘要
目的探讨微小RNA-101(miR-101)在子痫前期(PE)患者胎盘组织中的表达和作用及可能的调控机制。方法收集2012年1月至2015年12月咸宁市中心医院收治的40例PE患者和40例正常孕妇胎盘组织,采用荧光定量PCR分析胎盘组织中miR-101的表达情况。体外转染miR-101抑制物至人滋养层细胞系(HTR-8/SVneo),并观察其对细胞侵袭及凋亡的影响。通过生物信息学,双荧光素酶报告基因试验及蛋白质印迹分析miR-101靶基因。结果 miR-101在PE患者胎盘组织的表达量明显高于正常胎盘组织(P<0.05),而基质金属蛋白酶-1(MMP-1)在PE患者胎盘组织的表达量明显低于正常胎盘组织(P<0.05),二者表达水平呈明显负相关(P<0.05)。体外敲低miR-101可明显促进HTR-8/SVneo细胞的侵袭(P<0.05),并抑制细胞凋亡(P<0.05)。MMP-1是miR-101的靶基因,miR-101可通过与MMP-1信使RNA的3’-非编码区(3’-UTR)结合并抑制其翻译。体外敲低miR-101可显著增加MMP-1的蛋白表达水平(P<0.05)。结论 miR-101在PE患者胎盘组织中呈现高表达,敲低miR-101可显著促进胎盘细胞的侵袭并抑制其凋亡。MMP-1是miR-101的下游直接靶基因,提示miR-101可能通过负性调控MMP-1参与子痫前期的发生发展过程。
Objective To determine the expression and function of microRNA-101(miR-101)in placentas of preeclampsia(PE),and identify its potential regulatory mechanism.Methods Forty placentas of PE pregnancies and normal pregnancies were obtained at the Central Hospital of Xianning City between January 2012 and December 2015.Realtime PCR was performed to detect the expression levels of miR-101 in the placentas of PE pregnancies and normal pregnancies.The invasion and apoptosis of HTR-8/SVneo cells were detected after the cells were transfected with miR-101 inhibitors.Bioinformatic analysis,dual luciferase reporter assay,and Western blot analysis were used to identify target gene of miR-101 in HTR-8/SVneo cells.Results The miR-101 expression was significantly higher in placentas of PE pregnancies than in placentas of normal pregnancies(P<0.05),while the levels of MMP-1 mRNA was markedly lower in placentas of PE pregnancies than in placentas of normal pregnancies(P<0.05).MiR-101 expression was negatively correlated with MMP-1 mRNA in placentas of PE pregnancies(P<0.05).Knockdown of miR-101 significantly promoted invasion of HTR-8/SVneo cells(P<0.05),and suppressed cell apoptosis(P<0.05).MMP-1 was a target gene of miR-101 in HTR-8/SVneo cells;miR-101 repressed MMP-1 expression via binding to its m RNA 3’-untranslated regions(3’-UTR)(P<0.05).Knockdown of miR-101 in vitro significantly increased the protein expression level of MMP-1(P<0.05).Conclusion miR-101 is up-regulated in placentas of PE pregnancies;knockdown of miR-101 significantly promotes invasion of HTR-8/SVneo cells,and suppress cell apoptosis.MMP-1 is a direct target gene of miR-101 in HTR-8/SVneo cells,indicating miR-101 may be involved in the occurrence and development of PE by negatively regulating MMP-1 expression.
作者
毛雪宝
吕俊
MAO Xue-bao;LYU Jun(Department of Obstetrics and Gynecology,the Central Hospital of Xianning City,the Affiliated Hospital of Hubei College of Science and Technology,Xianning 437100,China)
出处
《中国实用妇科与产科杂志》
CAS
CSCD
北大核心
2018年第12期1393-1396,共4页
Chinese Journal of Practical Gynecology and Obstetrics
基金
国家自然科学基金青年基金(81200451)
