MiR-218对滋养层细胞迁移侵袭的影响及其机制研究

doi:10.12047/j.cjap.5453.2017.043

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

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. 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. 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/Abstract

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

陈育娟, 吴培阳, 高瑞秋. MiR-218对滋养层细胞迁移侵袭的影响及其机制研究[J]. 中国应用生理学杂志, 2017, 33(2): 169-173.

CHEN Yu-juan, WU Pei-yang, GAO Rui-qiu. MiR-218 inhibits HTR-8 cells migration and invasion by targeting SOX4[J]. CJAP, 2017, 33(2): 169-173.

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MiR-218对滋养层细胞迁移侵袭的影响及其机制研究

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

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