| 杨慧丽,杨俊娟,李新敏.微小 RNA-21调控同源性磷酸酶 -张力蛋白基因表达在子痫前期发病中的作用机制[J].安徽医药,2021,25(4):709-713. |
| 微小 RNA-21调控同源性磷酸酶 -张力蛋白基因表达在子痫前期发病中的作用机制 |
| The mechanism of microRNA-21 regulating phosphatase and tensin homolog expression in the pathogenesis of preeclampsia |
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| DOI：10.3969/j.issn.1009-6469.2021.04.018 |
| 中文关键词: 子痫前期 基因，肿瘤抑制 微小 RNA-21 同源性磷酸酶 -张力蛋白 HTR-8/SVneo细胞 |
| 英文关键词: Preeclampsia Genes, tumor suppressor microRNA-21 Phosphatase and tensin homolog HTR-8/SVneo cells |
| 基金项目:河南省医学科技攻关计划项目(2018020716) |
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| 中文摘要: |
| 目的探究微小 RNA-21(miR-21)在子痫前期(PE)病人发病中的表达变化及可能作用机制。方法收集 2016年 8月至 2019年 10月郑州市妇幼保健院收治的 45例 PE病人及 45例匹配正常孕妇胎盘组织，采用实时定量 PCR(RT-qPCR)法检测 miR21相对表达水平。体外培养人滋养层细胞系 HTR-8/SVneo，转染 miR-21模拟物(mimics组)，抑制物(inhibitor组)及无意义序列(NC组)，另设无任何处理 HTR-8/SVneo细胞为空白对照组(BC组)。采用 Annexin V-FITC/PI双染色流式细胞凋亡检测试剂盒检测细胞凋亡率，采用 Tanswell小室检测细胞侵袭性。生物学分析并进行双荧光素酶报告基因验证 miR-21与同源性磷酸酶 -张力蛋白基因(PTEN)的靶向关系，免疫印迹(WB)法检测胎盘组织及细胞中 PTEN、磷脂酰肌醇 -3激酶(p-PI3K)及蛋白激酶 B(p-Akt)磷酸化蛋白表达情况。结果与正常组(0.45±0.06)比较， PE病人胎盘组织中 miR-21表达水平(0.41±0.05)显著降低(t=79.157，P <0.05)PTEN mRNA及蛋白表达水平分别为(4.13±0.58)(0.91±0.08)均显著增加(t=36.201、30.858，P<0.05)。 PE病人胎盘组织中 miR-21，与 PTEN蛋白表达水平呈负相关(r=?0.542，P<0.0、5)。与 BC组、 NC组比较， mimics组 HTR-8/SVneo细胞侵袭细胞数、 pPI3K、p-Akt蛋白表达水平显著增加，凋亡率、 PTEN蛋白表达水平显著降低， inhibitor组呈相反趋势。双荧光素酶报告基因实验结果显示，与转染 NC+Wt(1.00±0.00)比较，转染 mimics+Wt后 HTR-8/SVneo细胞相对荧光活性(0.39±0.04)显著降低(t=37.355，P<0.05)。结论 miR-21可通过靶向抑制 PTEN表达促进细胞侵袭并抑制其凋亡，可能与促进 PI3K/Akt通路激活有关。 |
| 英文摘要: |
| Objective To explore the expression change of microRNA-21 (miR-21) in the pathogenesis of preeclampsia (PE) patients and its possible mechanism.Methods Placental tissues of forty-five PE patients treated in Maternal and Child Care Hospital of Zheng zhou City from August 2016 to October 2019 and 45 matched normal pregnant women were collected, and the relative expression level ofmiR-21 wasdetectedby real-timequantitativePCR (RT-qPCR).Human trophoblastHTR-8/SVneocelllinewas cultured invitroand transfected with mimic of miR-21 (mimics group), inhibitor (inhibitor group) and nonsense sequence (NC group), in addition, HTR-8/SVneo cells without any treatment were used as the blank control group (BC group). Annexin V-FITC/PI double staining flow cytometry was usedto detect the apoptosis rate, and Tanswell cell was used to detect cell invasion. The target relationship between miR-21 and phosphataseand tensin homolog (PTEN) was verified by biological analysis and double luciferase reporter gene. The expressions of PTEN, phosphatidylinositol-3 kinase (p-PI3K) and protein kinase B (p-Akt) phosphorylated protein in placental tissues and cells were detected by Western blot (WB).Results Compared with the normal group (0.45±0.06), the expression level of miR-21 in placenta of PE patients (0.41±0.05) decreased significantly (t=79.157, P<0.05), and the expression levels of PTEN mRNA and protein were (4.13±0.58), (0.91±0.08), which both increased significantly (t=36.201, 30.858, P < 0.05). There was a negative correlation between miR-21 and PTEN protein expression in placenta of PE patients (r=?0.542, P<0.05).Compared with BC group and NC group, the number of invasive cells in HTR-8/SVneo cells, p-PI3K and p-Akt protein expression levels in mimics group increased significantly, while the apoptosis rate and PTEN protein expressionlevel decreased significantly. The inhibitor group showed an opposite trend. The results of double luciferase reporter gene experimentshowed that the relative fluorescence activity of HTR-8/SVneo cells (0.39±0.04) was significantly decreased after transfection of mimics+Wt compared with the transfection of NC+Wt (1.00±0.00) (t=37.355, P<0.05).Conclusion miR-21 can promote cell invasion and inhibitapoptosisbytargetinginhibition of PTEN expression,whichmaybe related totheactivationofPI3K/Aktpathway. |
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