miR-141-3p 表达对人异位子宫内膜细胞增殖与凋亡影响及其机制(1)
[摘要] 目的 探讨微小RNA-141-3p(miR-141-3p)对人异位子宫内膜细胞(HEEC)增殖和凋亡的影响及其机制。
方法 采用实时荧光定量聚合酶链反应(qRT-PCR)检测异位子宫内膜组织及正常子宫内膜组织中miR-141-3p的表达。体外原代培养HEEC，miR-141-3p mimics转染至HEEC后，采用MTT法检测细胞活力，流式细胞术检测细胞凋亡率。采用双荧光素酶报告实验验证miR-141-3p与高迁移率族蛋白B1(HMGB1)的靶向调控作用。
结果 异位子宫内膜组织中miR-141-3p的表达水平明显低于正常子宫内膜组织(t=56.880，P<0.001);miR-141-3p过表达能显著降低HEEC增殖活力(t=10.972，P<0.001)，升高细胞凋亡率(t=18.233，P<0.001);miR-141-3p可靶向结合HMGB1。
结论 miR-141-3p过表达可通过靶向调控HMGB1而抑制HEEC增殖，诱导其凋亡。
[关键词] 子宫内膜异位症;微RNAs;靶基因修复;高迁移率族蛋白质类;细胞增殖;细胞凋亡
[中图分类号] R711.71;R394.3
[文献标志码] A
[文章编号] 2096-5532(2020)03-0342-05
doi：10.11712/jms.2096-5532.2020.56.108
[开放科学(资源服务)标识码(OSID)]
[网络出版] https：//kns.cnki.net/kcms/detail/37.1517.R.20200610.1435.007.html;2020-06-11 11：19
EFFECT OF MIR-141-3P EXPRESSION ON THE PROLIFERATION AND APOPTOSIS OF HUMAN ECTOPIC ENDOMETRIAL CELLS AND RELATED MECHANISM
LIU Wenbo， FENG Quanling
(Department of Obstetrics and Gynecology， The Third Affiliated Hospital of Zhengzhou University， Zhengzhou 450052， China)
[ABSTRACT]ObjectiveTo investigate the effect of miR-141-3p on the proliferation and apoptosis of human ectopic endometrial cells (HEECs) and related mechanism.
MethodsQuantitative real-time PCR was used to measure the expression of miR-141-3p in ectopic endometrial tissue and normal endometrial tissue. Primary HEECs were cultured in vitro， and after miR-141-3p mimics were transfected into HEECs， MTT assay was used to measure cell viability and flow cytometry was used to mea-
sure apoptosis rate. The dual-luciferase reporter experiment was used to verify the targeted regulatory effect of miR-141-3p and high-mobility group box 1 (HMGB1).
ResultsThe expression level of miR-141-3p in ectopic endometrial tissue was significantly lower than that in normal endometrial tissue (t=56.880，P<0.001). Overexpression of miR-141-3p can significantly reduce the proliferative activity of HEECs (t=10.972，P<0.001) and increase cell apoptosis rate (t=18.233，P<0.001). miR-141-3p could target HMGB1.
ConclusionmiR-141-3p overexpression may inhibit the proliferation of HEECs and induce apoptosis through targeted regulation of HMGB1.
[KEY WORDS]endometriosis; microRNAs; targeted gene repair; high mobility group proteins; cell proliferation; apoptosis
目前關于子宫内膜异位症发病机制尚未完全阐明[1-2]。既往研究已表明，体腔化生、血管转移等均可能引发子宫内膜异位症，而人异位子宫内膜细胞(HEEC)恶性增殖及凋亡能力降低是子宫内膜异位症的主要病理机制[3-4]。因此，深入探究HEEC增殖及凋亡的分子机制有助于提高子宫内膜异位症治疗效果。研究已表明，微小RNA-141(microRNA-141，miR-141)在子宫内膜异位症病人中表达下调，但关于其具体作用机制尚未明确[5-6]。miR-141是微小RNA-141-3p(microRNA-141-3p，miR-141-3p)的前体，关于miR-141-3p对HEEC生物行为的影响及其机制研究相对较少。已有研究表明，子宫内膜异位症病人中高迁移率族蛋白 B1(HMGB1)的表达水平升高，其可能通过调节新生血管生成而参与子宫内膜异位症发生过程[7]。Target Scan网站预测显示，HMGB1可能是miR-141-3p的靶基因，但miR-141-3p是否可通过靶向调控HMGB1而影响HEEC的增殖及凋亡尚未明确。为此，本研究探讨miR-141-3p表达对HEEC增殖及凋亡的影响，分析其与HMGB1的靶向调控关系及其可能作用机制。现将结果报告如下。, http://www.100md.com(刘文博 封全灵)
方法 采用实时荧光定量聚合酶链反应(qRT-PCR)检测异位子宫内膜组织及正常子宫内膜组织中miR-141-3p的表达。体外原代培养HEEC，miR-141-3p mimics转染至HEEC后，采用MTT法检测细胞活力，流式细胞术检测细胞凋亡率。采用双荧光素酶报告实验验证miR-141-3p与高迁移率族蛋白B1(HMGB1)的靶向调控作用。
结果 异位子宫内膜组织中miR-141-3p的表达水平明显低于正常子宫内膜组织(t=56.880，P<0.001);miR-141-3p过表达能显著降低HEEC增殖活力(t=10.972，P<0.001)，升高细胞凋亡率(t=18.233，P<0.001);miR-141-3p可靶向结合HMGB1。
结论 miR-141-3p过表达可通过靶向调控HMGB1而抑制HEEC增殖，诱导其凋亡。
[关键词] 子宫内膜异位症;微RNAs;靶基因修复;高迁移率族蛋白质类;细胞增殖;细胞凋亡
[中图分类号] R711.71;R394.3
[文献标志码] A
[文章编号] 2096-5532(2020)03-0342-05
doi：10.11712/jms.2096-5532.2020.56.108
[开放科学(资源服务)标识码(OSID)]
[网络出版] https：//kns.cnki.net/kcms/detail/37.1517.R.20200610.1435.007.html;2020-06-11 11：19
EFFECT OF MIR-141-3P EXPRESSION ON THE PROLIFERATION AND APOPTOSIS OF HUMAN ECTOPIC ENDOMETRIAL CELLS AND RELATED MECHANISM
LIU Wenbo， FENG Quanling
(Department of Obstetrics and Gynecology， The Third Affiliated Hospital of Zhengzhou University， Zhengzhou 450052， China)
[ABSTRACT]ObjectiveTo investigate the effect of miR-141-3p on the proliferation and apoptosis of human ectopic endometrial cells (HEECs) and related mechanism.
MethodsQuantitative real-time PCR was used to measure the expression of miR-141-3p in ectopic endometrial tissue and normal endometrial tissue. Primary HEECs were cultured in vitro， and after miR-141-3p mimics were transfected into HEECs， MTT assay was used to measure cell viability and flow cytometry was used to mea-
sure apoptosis rate. The dual-luciferase reporter experiment was used to verify the targeted regulatory effect of miR-141-3p and high-mobility group box 1 (HMGB1).
ResultsThe expression level of miR-141-3p in ectopic endometrial tissue was significantly lower than that in normal endometrial tissue (t=56.880，P<0.001). Overexpression of miR-141-3p can significantly reduce the proliferative activity of HEECs (t=10.972，P<0.001) and increase cell apoptosis rate (t=18.233，P<0.001). miR-141-3p could target HMGB1.
ConclusionmiR-141-3p overexpression may inhibit the proliferation of HEECs and induce apoptosis through targeted regulation of HMGB1.
[KEY WORDS]endometriosis; microRNAs; targeted gene repair; high mobility group proteins; cell proliferation; apoptosis
目前關于子宫内膜异位症发病机制尚未完全阐明[1-2]。既往研究已表明，体腔化生、血管转移等均可能引发子宫内膜异位症，而人异位子宫内膜细胞(HEEC)恶性增殖及凋亡能力降低是子宫内膜异位症的主要病理机制[3-4]。因此，深入探究HEEC增殖及凋亡的分子机制有助于提高子宫内膜异位症治疗效果。研究已表明，微小RNA-141(microRNA-141，miR-141)在子宫内膜异位症病人中表达下调，但关于其具体作用机制尚未明确[5-6]。miR-141是微小RNA-141-3p(microRNA-141-3p，miR-141-3p)的前体，关于miR-141-3p对HEEC生物行为的影响及其机制研究相对较少。已有研究表明，子宫内膜异位症病人中高迁移率族蛋白 B1(HMGB1)的表达水平升高，其可能通过调节新生血管生成而参与子宫内膜异位症发生过程[7]。Target Scan网站预测显示，HMGB1可能是miR-141-3p的靶基因，但miR-141-3p是否可通过靶向调控HMGB1而影响HEEC的增殖及凋亡尚未明确。为此，本研究探讨miR-141-3p表达对HEEC增殖及凋亡的影响，分析其与HMGB1的靶向调控关系及其可能作用机制。现将结果报告如下。, http://www.100md.com(刘文博 封全灵)