lncRNA HAND2-AS1通过调节miR-21表达对子宫内膜异位症患者子宫内膜基质细胞迁移和侵袭的抑制作用  

Inhibitory effect of lncRNA HAND2-AS1 on migration and invasion of endometrial stromal cells in patients with endometriosis by regulating expression of miR-21

作　　者：苗卉[1] 苗聪秀[1] 李娜[1] 韩晶[1] MIAO Hui;MIAO Congxiu;LI Na;HAN Jing(Department of Reproductive Genetics,Heping Hospital,Changzhi Medical College,Institute of Reproduction and Genetics of Changzhi Medical College,Key Labrotory of Reproduction and Genetics of Changzhi Medical College,Key Labrotory of Reproduction Engineer of Shanxi Health Committee,Changzhi 046000,China)

机构地区：[1]长治医学院附属和平医院生殖遗传科长治医学院生殖与遗传研究所长治医学院生殖与遗传重点实验室山西省卫健委生殖工程重点实验室,山西长治046000

出　　处：《吉林大学学报（医学版）》2023年第3期733-741,共9页Journal of Jilin University：Medicine Edition

基　　金：国家自然科学基金项目(31971073);山西省科技厅应用基础研究计划项目(201901D111325);山西省教育厅科技创新项目(2020L0383)。

摘　　要：目的:探讨子宫内膜异位症(EMT)患者子宫内膜组织中长链非编码RNAs(lncRNAs)HAND2-AS1对异位子宫内膜(EC)组织中子宫内膜基质细胞(ESCs)增殖、迁移和侵袭能力的影响,并阐明其可能机制。方法:收集30例EMT患者(EMT组)的EC组织和30例健康育龄女性(对照组)的子宫内膜组织,分离子宫内膜组织获取ESCs。采用脂质体转染法转染EMT患者EC组织的ESCs,并将ESCs分为pcDNA组(转染pcDNA空质粒)、HAND2-AS1组(转染HAND2-AS1过表达质粒)、mimic NC组(转染mimic NC)、miR-21 mimic组(转染miR-21 mimic)、pcDNA+mimic NC组(联合转染pcDNA和mimic NC)、HAND2-AS1+mimic NC组(联合转染HAND2-AS1过表达质粒和mimic NC)、pcDNA+miR-21 mimic组(联合转染pcDNA空质粒和miR-21 mimic)和HAND2-AS1+miR-21 mimic组(联合转染HAND2-AS1过表达质粒和miR-21 mimic),另设未转染的细胞为空白对照组。采用实时荧光定量PCR(RT-qPCR)法检测2组研究对象子宫内膜组织和ESCs中HAND2-AS1 mRNA和miR-21表达水平,采用Pearson相关系数分析其相关性。采用生物信息学软件Starbase预测lncRNA HAND2-AS1与miR-21的靶向作用关系,荧光素酶基因报告实验进行验证。CCK-8法检测各组ESCs增殖活性,Transwell小室实验检测ESCs的迁移和侵袭数。结果:2组研究对象年龄、体质量指数(BMI)和血清中卵泡刺激素(FSH)、黄体生成素(LH)及催乳素(PRL)水平比较差异均无统计学意义(P<0.05)。与对照组比较,EMT组患者血清中雌二醇(E2)水平升高(P<0.05),EMT组患者EC组织中HAND2-AS1 mRNA表达水平降低(P<0.05),miR-21表达水平升高(P<0.05);与对照组比较,EMT组ESCs中HAND2-AS1 mRNA表达水平降低(P<0.05),miR-21表达水平明显升高(P<0.01)。Pearson相关系数分析,对照组研究对象HAND2-AS1与miR-21表达水平无明显相关性(r=0.34,P>0.05),EMT组患者ES组织中和ESCs中HAND2-AS1与miR-21表达水平呈负相关关系(r=-0.57,P<0.05)。RT-qPCR法检测,与空白对照组比较,HAND2-AS1组ESCs中HAND2-AS1 mRNObjective:To discuss the effect of long non-coding RNA(lncRNA)HAND2-AS1 in endometrium tissue of the patients with endometriosis(EMT)on the proliferation,migration and invasion of the endometrial stromal cells(ESCs)in ectopic endometrium(EC)tissue,and to clarify the possible mechanism.Methods:The EC tissue of 30 patients with EMT(EMT group)and the endometrium tissue of 30 healthy women of childbearing age(control group)were collected;the endometrium tissue was isolated,and the ESCs were collected.The ESCs in EC tissue of the EMT patients were transfected with liposome transfection method,and the ESCs were divided into pcDNA group(transfected with pcDNA empty plasmid),HAND2-AS1 group(transfected with HAND2-AS1 over-expression plasmid),mimic NC group(transfected with mimic NC),miR-21 mimic group(transfected with miR-21 mimic),pcDNA+mimic-NC group(transfected with pcDNA and mimic NC),HAND2-AS1+mimic NC group(transfected with HAND2-AS1 over-expression plasmid and mimic NC),pcDNA+miR-21 mimic group(transfected with pcDNA empty plasmid and miR-21 mimic),and HAND2-AS1+miR-21 mimic group(transfected with HAND2 AS1 over-expression plasmid and miR-21 mimic),and the cells without any transfection were used as blank control group.The expression levels of HAND2-AS1 mRNA and miR-21 in the endometrium tissue and ESCs of the subjects in two groups were detected by real-time fluorescence quantitative PCR(RT-qPCR)method;and the relationship between them was analyzed by Pearson correlation coefficient;bioinformatics software Starbase was used to predict the targeting relationship between lncRNA HAND2-AS1 and miR-21;the luciferase gene reporting assay was used to verify.The proliferation activities of the ESCs in various groups were detected by CCK-8 method;the numbers of migration and invasion cells were detected by Transwell chamber assay.Results:There were no significant differences in the age,body mass index(BMI),serum levels of follicle stimulating hormone(FSH),luteinizing hormone(LH)and prolactin(PRL)of the subjects between tw

关 键 词：子宫内膜异位症 异位子宫内膜 长链非编码RNA HAND2-AS1 MIR-21 子宫内膜基质细胞 细胞迁移 细胞侵袭 

分 类 号：R711.2[医药卫生—妇产科学]