miR-30b在子宫内膜异位症中的表达及功能研究

Study on the expression and function of miR-30b in endometriosis

目的检测miR-30b在子宫内膜异位症中的表达水平,并探究miR-30b对子宫内膜异位症的治疗作用。方法2018年7月至2020年7月在温州医科大学附属第二医院手术获取的27例子宫内膜异位症患者病理组织为子宫内膜异位症组,15例子宫肌瘤患者子宫内膜组织为正常子宫内膜组。实时定量PCR(qRT-PCR)检测组织中miR-30b的表达量。培养人原代子宫内膜间质细胞(HESCs),5-乙炔基-2′-脱氧尿苷掺入(EdU)法来检测miR-30b对细胞增殖能力的影响。根据starBase 2.0软件预测与miR-30b相互作用的长链非编码RNA(lncRNA),使用qRT-PCR初步验证该lncRNA在异位内膜中的表达量及与miR-30b的相互关系。进一步建造子宫内膜异位症SD大鼠模型,在活体实验水平,使用miR-30b的模拟物来验证miR-30b对子宫内膜异位症异位囊肿的治疗作用。结果与正常子宫内膜(EN)相比,miR-30b在子宫内膜异位症(EC)中的表达显著下降。miR-30b的外源性过表达抑制了HESCs的增殖能力,同时,下调miR-30b的表达量可促进HESCs的增殖能力。StarBase 2.0软件预测lncRNA OIP5-AS1与miR-30b有多个结合位点,与正常内膜比较,lncRNA OIP5-AS1在异位内膜中表达上调,且与miR-30b的表达量呈负相关。此外,在子宫内膜异位症的大鼠模型中,miR-30b模拟物显著抑制子宫内膜异位症异位囊肿的生长。结论miR-30b在异位子宫内膜中低表达且抑制HESCs的增殖能力,可能成为子宫内膜异位症潜在的治疗靶点。

Objective To detect the expression level of miR-30b in endometriosis and explore the therapeutic effect of miR-30b on endometriosis.Methods All specimens were obtained during surgery in the Second Affiliated Hospital of Wenzhou Medical University from July 2018 to July 2020.Quantitative real-time PCR(qRT-PCR)was used to detect the expression of miR-30b in tissues.The pathological tissues of 27 patients with endometriosis were extracted as the endometriosis group,and the endometrial tissues of 15 patients with uterine fibroids were extracted as the normal endometrium group.Human primary endometrial stromal cells(HESCs)were cultured,and 5-ethynyl-2′-deoxyuridine incorporation(EdU)assay was used to detect the effect of miR-30b on cell proliferation ability.Long non-coding RNAs(lncRNAs)that interact with miR-30b were predicted according to starBase 2.0 software.The expression of this lncRNA in ectopic endometrium and its interrelationship with miR-30b were preliminarily validated using qRT-PCR.Furthermore,a SD rat model of endometriosis was constructed.At the in vivo level,miR-30b mimics were used to validate the therapeutic effect of miR-30b on endometriotic ectopic cysts.Results The expression of miR-30b was significantly decreased in ectopic endometrium(EC)compared with normal endometrium(EN).Exogenous overexpression of miR-30b inhibited the proliferation ability of HESCs.Meanwhile,down-regulation of miR-30b expression promoted the proliferation ability of HESCs.StarBase 2.0 software predicted that lncRNA OIP5-AS1 had multiple binding sites with miR-30b,and lncRNAOIP5-AS1 was up-regulated in ectopic endometrium compared with normal endometrium,and was negatively correlated with the expression of miR-30b.In addition,in a rat model of endometriosis,we observed that miR-30b mimics significantly inhibited the growth of endometriotic ectopic cysts.Conclusion miR-30b is lowly expressed in ectopic endometrium and inhibits the proliferation of HESCs,which may become a potential therapeutic target for endometriosis.