摘要
目的 研究微小RNA?17?5p(miR?17?5p)对子宫内膜异位症(EMs)在位子宫内膜间质细胞(ESC)增殖和凋亡的影响.方法 选择2016年2月~2017年6月本院收治的EMs患者30例(EMs组),同期因卵巢囊肿或浆膜下肌瘤等行手术治疗的患者30例作为对照组,分离、原代培养两组ESC,应用噻唑蓝(MTT)比色法、流式细胞仪分别检测细胞的增殖和凋亡,双萤光素酶报告系统验证miR?17?5p和S100钙结合蛋白A4(S100A4)的关系,实时荧光定量PCR(qRT?PCR)检测miR?17?5p和S100A4 RNA的表达,免疫印迹法检测S100A4蛋白表达水平.结果 与对照组相比,EMs组中miR?17?5p表达水平下降(0.53±0.05比1.01±0.08,P<0.05),S100A4 mRNA和蛋白的表达水平上升(5.27±0.25比1.03±0.09、0.59±0.05比0.23±0.03,均P<0.05);双萤光素酶报告系统结果显示,miR?17?5p靶向负调控S100A4的表达;过表达miR?17?5p和抑制S100A4均可抑制EMs ESC细胞的增殖活性,诱导细胞凋亡;过表达S100A4可逆转上调miR?17?5p对EMs ESC细胞增殖和凋亡的影响.结论 miR?17?5p通过靶向S100A4调控EMs在位内膜细胞的增殖和凋亡.
Objective To investigate the effects of microRNA?17?5p(miR?17?5p)on proliferation and apoptosis of eutopic endometrial stromal cells(ESC)in endometriosis(EMs).Method A total of 30 EMs patients admitted to our hospital between February 2016 and June 2017(EMs group)were enrolled.A contemporary cohort of 30 patients with ovarian cysts or subserosal fibroids undergoing surgery in our hospital were set as the control group.ESC were isolated from the two groups and subject to primary culture.The proliferation and apoptosis of the ESCs were detected by MTT colorimetry and flow cytometry.The relationship between miR?17?5p and S100A4 was verified by dual luciferase reporter system.Real?time PCR(qRT?PCR)was used to examine the RNA expression levels of miR?17?5p and S100 calcium binding protein A4(S100A4)RNA,and the protein expression level of S100A4 was detected by immunoblotting.Results Compared with the control group,the expression level of miR?17?5p in the EMs group was lowered(0.53±0.05 vs 1.01±0.08,P<0.05),and the mRNA and protein expression levels of S100A4 was increased(5.27±0.25 vs 1.03±0.09,0.59±0.05 vs 0.23±0.03,both P<0.05).The results of dual luciferase reporter system showed that miR?17?5p negatively target?regulated the expression of S100A4.Overexpression of miR?17?5p and suppressed S100A4 expression were shown to inhibit the proliferation and induce apoptosis of ESC in EMs.Overexpression of S100A4 was shown to reversely up?regulate the effects of miR?17?5p on proliferation and apoptosis of ESC in EMs.Conclusion miR?17?5p may regulate the proliferation and apoptosis of eutopic ESC in EMs by targeting S100A4.
作者
冯双苗
袁银花
张化莲
Feng Shuangmiao;Yuan Yinhua;Zhang Hualian(No.2 Department of Obstetrics,Zhumadian Central Hospital,Zhumadian,Henan 463000,China)
出处
《中华生物医学工程杂志》
CAS
2019年第2期183-188,共6页
Chinese Journal of Biomedical Engineering