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中国应用生理学杂志 ›› 2017, Vol. 33 ›› Issue (2): 169-173.doi: 10.12047/j.cjap.5453.2017.043

• 研究论文 • 上一篇    下一篇

MiR-218对滋养层细胞迁移侵袭的影响及其机制研究

陈育娟1, 吴培阳2, 高瑞秋1   

  1. 1. 厦门市妇幼保健院产科, 福建 厦门 361000;
    2. 厦门大学附属第一医院医务科, 福建 厦门 361000
  • 收稿日期:2016-05-05 修回日期:2016-12-09 出版日期:2017-03-28 发布日期:2018-06-20
  • 通讯作者: 陈育娟,Tel:13950092828,E-mail:chenyujuanfzxm@sina.com E-mail:chenyujuanfzxm@sina.com
  • 基金资助:
    国家自然科学青年基金(U1204801)

MiR-218 inhibits HTR-8 cells migration and invasion by targeting SOX4

CHEN Yu-juan1, WU Pei-yang2, GAO Rui-qiu1   

  1. 1. Department of Obstetrics, Maternal and Child Hospital of Xiamen City, Xiamen 361000;
    2. Department of Medical, the First Affiliated Hospital of Xiamen University, Xiamen 361000, China
  • Received:2016-05-05 Revised:2016-12-09 Online:2017-03-28 Published:2018-06-20
  • Supported by:
    国家自然科学青年基金(U1204801)

摘要: 目的:研究miR-218是否通过下调SOX4影响滋养层细胞系HTR-8细胞的迁移和侵袭。方法:妊娠期高血压疾病(HDCP)患者46例,平均年龄(31 ±4.6)岁,收缩期血压≥ 140 mmHg和/或舒张期血压> 90 mmHg;以血压正常孕妇50例为对照,实时荧光定量PCR(RT-PCR)检测两组患者静脉血中miR-218的表达情况。转染miR-218mimic和miR-NC至离体培养的HTR-8细胞中,将细胞分为对照组(加入DMEM)、空质粒组(加入miR-NC)和过表达miR-218组(加入miR-218 mimic)3组,检测细胞的迁移侵袭情况以及细胞中MMP-2和MMP-9的表达,,生物信息学预测miR-218潜在靶基因为SOX4,利用荧光素酶素试验验证SOX4是miR-218的靶基因;再通过转染过表达SOX4的质粒至HTR-8细胞,HTR-8细胞分为过表达miR-218组、过表达miR-218+空质粒组、过表达miR-218+SOX4组,以上方法检测HTR-8细胞的迁移侵袭情况。结果:相比于正常孕妇组,HDCP组患者血清中miR-218表达减少(P <0.01)。相比于空质粒组,转染miR-218mimic后,HTR-8细胞中MMP-2、MMP-9、SOX4的表达减少(P < 0.01),细胞迁移和侵袭能力下降(P < 0.01);荧光素酶试验结果显示,miR-218能够显著降低SOX4-3'-UTR质粒的荧光素活性(P< 0.01);相比于miR-218+空质粒组,转染过表达SOX4质粒后,HTR-8细胞迁移和侵袭能力增加(P < 0.01)。结论:HDCP患者血清中miR-218表达减少,miR-218可以通过下调SOX4从而抑制HTR-8细胞的迁移和侵袭。

关键词: 妊娠期高血压疾病, 微小RNA218, SOX4, 滋养层细胞, 侵袭

Abstract: Objective: To verify whether miR-218 could inhibit human trophoblastic cell (HTR-8 cells) migration and invasion by target-ing sex determining region Y-box 4(SOX4). Methods: The serum samples were collected from 46 hypertensive disorder complicating pregnan-cy (HDCP) and 50 normal pregnant women. RT-PCR was used to test the expression of miR-218 in the serum. In vitro, MiR-218 was trans-fe cted into HTR-8 cells. The HTR-8 cells were divided into three groups:normal control group, mimic control and miR-218 mimic group. The migratory and invasion ability of HTR-8 cells was tested, and the expressions of matrix metalloproteinase-2(MMP-2), MMP-9 and Sox4 were also investigated in the cells of each group. Luciferase assay was used to confirme whether Sox4-3'-UTR was the target gene of miR-218. Results: The expression of miR-218 was decreased in the serum of HDCP patients compared with the normal pregnant woman(P < 0.01). In vit-ro, compared with the control group, the invasion and migration ability of HTR-8 cells and the expression of MMP-2 MMP-9 and SOX4 were decreased in the miR-218 group (P < 0.01); The Luciferase activity of the SOX4-3'-UTR plasmid was significantly suppressed by miR-218 (P < 0.01); Over expression of SOX4 could reverse the effect of miR-218 on HTR-8 cells(P < 0.01). Conclusion: The expression of miR-218 decreases in the serum of HDPC patients and miR-218 inhibits HTR-8 cells invasion by targeting SOX4-3'-UTR.

Key words: hypertensive disorder complicating pregnancy (HDCP), miR-218, sex determining region Y-box 4, trophoblast cell, invasion

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