天津医药

天津医药 ›› 2021, Vol. 49 ›› Issue (10): 1037-1042.doi: 10.11958/20211062

• 细胞与分子生物学 • 上一篇    下一篇

miR-34a靶向MMP2对TNF-α诱导人绒毛膜滋养层细胞 HTR-8/SVneo迁移及侵袭的影响

温彦静，彭青，王静娜，李曼，常美英   

  1. 1河北省中医院产一科（邮编050032）；2定州市中医医院妇产科
  • 收稿日期:2021-04-30 修回日期:2021-06-14 出版日期:2021-10-15 发布日期:2021-10-15
  • 通讯作者: 温彦静 E-mail:wenyanjing852@163.com
  • 基金资助:
    河北省医学科学研究课题计划项目

Effects of miR-34a targeting MMP2 on migration and invasion of human villous trophoblasts HTR-8/SVneo induced by TNF-α

WEN Yan-jing, PENG Qing, WANG Jing-na, LI Man, CHANG Mei-ying   

  1. 1 Department of Obstetrics 1, Hebei Hospital of Traditional Chinese Medicine, Shijiazhuang 050032, China; 2 Department of
    Obstetrics and Gynecology, Dingzhou Hospital of Traditional Chinese Medicine
  • Received:2021-04-30 Revised:2021-06-14 Published:2021-10-15 Online:2021-10-15

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摘要： 目的 探讨微小RNA（miR）-34a靶向基质金属蛋白酶（MMP）2对肿瘤坏死因子（TNF）-α诱导人绒毛膜滋 养层细胞 HTR-8/SVneo 迁移和侵袭的影响。方法 体外培养人绒毛膜滋养层细胞 HTR-8/SVneo，利用 0.5 μg/L TNF-α刺激细胞；将细胞分为对照组、TNF-α诱导组、NC组、miR-34a mimics组、iNC组、miR-34a inhibitor组；实时荧 光定量PCR（qPCR）检测各组细胞中miR-34a、MMP2 mRNA的表达；CCK-8法、Transwell法和划痕实验分别检测各组 细胞增殖、侵袭和迁移情况；蛋白免疫印迹法检测MMP2蛋白的表达；双荧光素酶报告实验证明miR-34a和MMP2的 靶向关系。结果 与对照组比较，TNF-α 诱导组细胞 miR-34a 表达水平、细胞增殖抑制率显著升高（P＜0.05）， MMP2 mRNA和蛋白表达水平、细胞侵袭数目、细胞迁移率显著降低（P＜0.05）；与TNF-α诱导组和NC组比较，miR- 34a mimics组细胞miR-34a表达水平、细胞增殖抑制率显著升高（P＜0.05），MMP2 mRNA和蛋白表达水平、细胞侵袭 数目、细胞迁移率显著降低（P＜0.05）；与TNF-α诱导组和iNC组比较，miR-34a inhibitor组细胞miR-34a表达水平、 细胞增殖抑制率显著降低（P＜0.05），MMP2 mRNA和蛋白表达水平、细胞侵袭数目、细胞迁移率显著升高（P＜0.05）； miR-34a与MMP2具有靶向关系。结论 miR-34a可能通过负调控MMP2的表达抑制TNF-α诱导的人绒毛膜滋养 层细胞HTR-8/SVneo迁移和侵袭。

关键词: 微RNAs, 基质金属蛋白酶2, 肿瘤坏死因子α, 人绒毛膜滋养层细胞, 子痫前期, miR-34a

Abstract: Objective To investigate the effects of microRNA-34a (miR-34a) targeting matrix metalloproteinase 2 (MMP2) on the migration and invasion of human villous trophoblasts HTR-8/SVneo induced by tumor necrosis factor- α (TNF-α). Methods Human villous trophoblasts HTR-8/SVneo cells were cultured in vitro and stimulated with 0.5 μg/L TNF-α. Cells were divided into the control group, the TNF-α induction group, the NC group, the miR-34a mimics group, the iNC group and the miR-34a inhibitor group. The expression levels of miR-34a and MMP2 mRNA were detected by quantitative real time-PCR (qRT-PCR). The cell proliferation, invasion and migration were detected by cell counting kit 8 (CCK-8), transwell assay and scratch test. The expression of MMP2 protein was detected by Western blot assay. Luciferase reporter assay was used to prove the targeting relationship between miR-34a and MMP2. Results Compared with the control group, the expression level of miR-34a and the inhibition rate of cell proliferation were significantly increased in the TNF-α induction group (P＜0.05), and the expression levels of MMP2 mRNA and protein, the cell invasion number and the cell migration rate were significantly decreased (P＜0.05). Compared with the TNF-α induction group and NC group, the expression levels of miR-34a and the inhibition rate of cell proliferation were significantly increased in the miR-34a mimics group (P＜0.05), the expression levels of MMP2 mRNA and protein, the number of cell invasion and the cell migration rate were significantly decreased (P＜0.05). Compared with the TNF-α induction group and iNC group, the expression level of miR-34a and the inhibition rate of cell proliferation were significantly decreased in the miR-34a inhibitor group (P＜0.05), and the expression levels of MMP2 mRNA and protein, the number of cell invasion and the cell migration rate were significantly increased (P＜0.05). There was a targeting relationship between miR-34a and MMP2. Conclusion miR-34a may inhibit the invasion and migration of TNF-α induced human villous trophoblasts HTR-8/SVneo by negatively regulating the expression of MMP2.

Key words: microRNAs, matrix metalloproteinase 2, tumor necrosis factor-alpha, human villous trophoblasts, preeclampsia, miR-34a