摘要
目的:研究miR218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 mm Hg和/或舒张期血压>90 mm Hg;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miR218的表达情况。转染miR218mimic和miRNC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miRNC)和过表达miR218组(加入miR218 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miR218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miR218组、过表达miR218+空质粒组、过表达miR218+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miR218表达减少(P<0.01)。相比于空质粒组,转染miR218mimic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,miR218能够显著降低SOX4-3'-UTR质粒的荧光素活性(P<0.01);相比于miR218+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miR218表达减少,miR218可以通过下调SOX4从而抑制HTR-8细胞的迁移和侵袭。
Objective： To verify whether miR-218 could inhibit human trophoblastic cell （HTR-8 ceils） 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 transfected 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 ceils 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 vitro, 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 ceils（ 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.
出处
《中国应用生理学杂志》
CAS
CSCD
2017年第2期169-173,I0001,共6页
Chinese Journal of Applied Physiology
基金
国家自然科学青年基金(U1204801)
关键词
妊娠期高血压疾病
微小RNA218
SOX4
滋养层细胞
侵袭
hypertensive disorder complicating pregnancy（HDCP）
miR-218
sex determining region Y-box 4
trophoblast cell
invasion
