Effects of miR-375 knockdown on the proliferation and apoptosis of trophoblast cells in a preeclampsia model and its mechanism

CUI Xin; LI Qiu-hong; ZHANG Xiao-xue

doi:10.11659/jjssx.02E024023

您当前的位置：

首页 >

文章列表页 >

Effects of miR-375 knockdown on the proliferation and apoptosis of trophoblast cells in a preeclampsia model and its mechanism

Basic Research | 更新时间：2024-11-18

- Effects of miR-375 knockdown on the proliferation and apoptosis of trophoblast cells in a preeclampsia model and its mechanism
- JOURNAL OF REGIONAL ANATOMY AND OPERATIVE SURGERY Vol. 33, Issue 11, Pages: 980-984(2024)
- 作者机构：
  
  秦皇岛市妇幼保健院产科，河北秦皇岛 066000
- 作者简介：
- 基金信息：
- DOI：10.11659/jjssx.02E024023
  CLC： R714.24+4
- Published：25 November 2024，
  
  Received：15 February 2024，
- 稿件说明：
移动端阅览
崔鑫,李秋红,张小学.敲低miR-375对子痫前期模型滋养细胞增殖、凋亡的影响及机制[J].局解手术学杂志,2024,33(11):980-984.

CUI Xin,LI Qiu-hong,ZHANG Xiao-xue.Effects of miR-375 knockdown on the proliferation and apoptosis of trophoblast cells in a preeclampsia model and its mechanism[J].JOURNAL OF REGIONAL ANATOMY AND OPERATIVE SURGERY,2024,33(11):980-984.
崔鑫,李秋红,张小学.敲低miR-375对子痫前期模型滋养细胞增殖、凋亡的影响及机制[J].局解手术学杂志,2024,33(11):980-984. DOI： 10.11659/jjssx.02E024023.

CUI Xin,LI Qiu-hong,ZHANG Xiao-xue.Effects of miR-375 knockdown on the proliferation and apoptosis of trophoblast cells in a preeclampsia model and its mechanism[J].JOURNAL OF REGIONAL ANATOMY AND OPERATIVE SURGERY,2024,33(11):980-984. DOI： 10.11659/jjssx.02E024023.

摘要

目的

探讨敲低miR-375对子痫前期（PE）滋养细胞模型增殖、凋亡的影响及作用机制。

方法

取人绒毛膜滋养细胞株HTR-8/Svneo，随机分为常氧对照组、PE模型组、miR-NC组和miR-375 inhibitor组。其中，miR-NC组和miR-375 inhibitor组细胞分别转染miR-NC和miR-375 inhibitor；常氧对照组细胞在常氧环境下培养，其余3组细胞均在缺氧条件下培养以建立PE滋养细胞模型。采用RT-PCR检测miR-375的表达，EdU荧光染色、CCK-8实验检测细胞增殖情况，流式细胞术检测细胞凋亡情况，Western blot检测YAP1蛋白及凋亡相关蛋白（Bcl-2、caspase-3及Bax蛋白）的表达。双荧光素酶基因实验检测miR-375与YAP1的靶向调控关系。

结果

与常氧对照组比较，PE模型组细胞miR-375表达升高（

0.05），EdU阳性细胞百分比下降（

0.05），细胞增殖能力降低（

0.05），细胞凋亡率升高（

0.05），Bax、caspase-3蛋白表达水平升高（

0.05），YAP1、Bcl-2蛋白表达水平降低（

0.05）。PE模型组和miR-NC组细胞上述各项指标比较差异均无统计学意义（

0.05）。与miR-NC组比较，miR-375 inhibitor组细胞miR-375表达降低（

0.05），EdU阳性细胞百分比升高（

0.05），细胞增殖能力增强（

0.05），细胞凋亡率降低（

0.05），Bax、caspase-3蛋白表达降低（

0.05），YAP1、Bcl-2蛋白表达升高（

0.05）。双荧光素酶报告基因实验显示，miR-375对YAP1具有靶向调控作用（

0.05）。

结论

敲低miR-375可促进PE滋养细胞增殖并抑制细胞凋亡，其作用机制可能与其对YAP1基因的靶向调控有关。

Abstract

Objective

To investigate the effects of miR-375 knockdown on the proliferation and apoptosis of trophoblast cells in preeclampsia （PE） model and its mechanism.

Methods

Human chorionic trophoblast cell line HTR-8/Svneo was randomly divided into the normoxic control group， the PE model group， the miR-NC group， and the miR-375 inhibitor group. Among them， cells in the miR-NC group and miR-375 inhibitor group were transfected with miR-NC and miR-375 inhibitor， respectively；cells in the normoxic control group were cultured in a normoxic enviroment，and cells in the remaining three groups were cultured under hypoxic conditions to establish the PE trophoblast cell model. The expression of miR-375 was detected by RT-PCR， the proliferation of cells was detected by EdU fluorescence staining and CCK-8 assay， the apoptosis of cells was detected by flow cytometry， and the expression of YAP1 protein and apoptosis-related proteins （Bcl-2， caspase-3， and Bax proteins） were detected by Western blot. The target regulation relationship between miR-375 and YAP1 was detected by dual luciferase reporter gene assay.

Results

Compared with the normoxic control group， the PE model group showed an increase in the expression of miR-375 （

0.05）， a decrease in the proportion of EdU-positive cells （

0.05）， a decline in the cell proliferation ability （

0.05）， increases in the apoptosis rate （

0.05） and the expression levels of Bax and caspase-3 proteins （

0.05）， and a decrease in the expression levels of YAP1 and Bcl-2 proteins （

0.05）. There was no significant difference in the above indicators between the PE model group and the miR-NC group （

0.05）. Compared with the miR-NC group， the miR-375 inhibitor group showed a lower expression of miR-375 （

0.05）， an increased percentage of EdU-positive cells （

0.05）， an enhanced cell proliferation ability （

0.05）， a lower apoptosis rate （

0.05）， and a lower expression of Bax and caspase-3 proteins （

0.05）， while with a higher expression of YAP1 and Bcl-2 proteins （

0.05）. The dual luciferase reporter gene assay showed that miR-375 had a targeted regulatory effect on YAP1 （

0.05）.

Conclusion

Knocking down miR-375 can promote the proliferation and inhibit the apoptosis of PE trophoblast cells， and its mechanism may be related to its targeted regulation of YAP1 gene.

关键词

子痫前期miR-375YAP1细胞增殖细胞凋亡

Keywords

preeclampsiamiR-375YAP1cell proliferationcell apoptosis

references

Mol BWJ， Roberts CT， Thangaratinam S， et al. Pre-eclampsia［J］. Lancet， 2016， 387（10022）：999-1011. doi： 10.1016/S0140-6736（15）00070-7http://dx.doi.org/10.1016/S0140-6736（15）00070-7.

Jung E， Romero R， Yeo L， et al. The etiology of preeclampsia［J］. Am J Obstet Gynecol， 2022， 226（2S）：S844-S866. doi：10.1016/j.ajog.2021.11.1356http://dx.doi.org/10.1016/j.ajog.2021.11.1356.

Bakrania BA， Spradley FT， Drummond HA， et al. Preeclampsia： linking placental ischemia with maternal endothelial and vascular dysfunction［J］. Compr Physiol， 2020， 11（1）：1315-1349.doi：10.1002/cphy.c200008http://dx.doi.org/10.1002/cphy.c200008.

Liu S， Xie X， Lei H， et al. Identification of key circRNAs/lncRNAs/miRNAs/mRNAs and pathways in preeclampsia using bioinformatics analysis［J］. Med Sci Monit， 2019， 25：1679-1693.doi：10.12659/MSM.912801http://dx.doi.org/10.12659/MSM.912801.

Hromadnikova I，Kotlabova K，Dvorakova L， et al.Postpartum profiling of microRNAs involved in pathogenesis of cardiovascular/cerebrovascular diseases in women exposed to pregnancy-related complications［J］. Int J Cardiol， 2019， 291：158-167.doi：10.1016/j.ijcard.2019.05.036http://dx.doi.org/10.1016/j.ijcard.2019.05.036.

Liu Y， Wang Q， Wen J， et al. MiR-375： a novel multifunctional regulator［J］.Life Sci，2021，275（1）：119323.doi：10.1016/j.lfs.2021.119323http://dx.doi.org/10.1016/j.lfs.2021.119323.

Akgör U， Ayaz L， Çayan F. Expression levels of maternal plasma microRNAs in preeclamptic pregnancies［J］. J Obstet Gynaecol， 2021， 41（6）：910-914. doi：10.1080/01443615.2020.1820465http://dx.doi.org/10.1080/01443615.2020.1820465.

贾清淼，马丽丽，闫凤梅.早发型子痫前期患者血清miR-152，miR-375水平与妊娠结局关系［J］.中国计划生育学杂志，2023，31（8）：1943-1947.doi：10.3969/j.issn.1004-8189.2023.08.037http://dx.doi.org/10.3969/j.issn.1004-8189.2023.08.037.

Ren Y， Xu Y， Wang Y， et al. Regulation of miR-375 and Sonic hedgehog on vascular endothelial growth factor in preeclampsia rats and its effect on trophoblastcells［J］. Biosci Rep， 2020， 15（1）：BSR20200613. doi：10.1042/BSR20200613http://dx.doi.org/10.1042/BSR20200613.

Raghupathy R. Cytokines as key players in the pathophysiology of preeclampsia［J］. Med Princ Pract， 2013， 22（Suppl 1）：8-19.doi：10.1159/000354200http://dx.doi.org/10.1159/000354200.

张玉月，朱亚飞，高丽娜，等.环状RNA BRAF_2在子痫前期胎盘滋养细胞增殖和凋亡中的作用及其机制［J］.解放军医学杂志，2023，48（4）：374-382. doi：10.11855/j.issn.0577-7402.2023.04.0374http://dx.doi.org/10.11855/j.issn.0577-7402.2023.04.0374.

杨晓涛，韩顶立，Philip N.Baker，等.体外缺氧诱导子痫前期滋养细胞模型优化及代谢组学鉴定［J］.重庆医科大学学报，2017，42（8）：1045-1052.doi：10.13406/j.cnki.cyxb.001343http://dx.doi.org/10.13406/j.cnki.cyxb.001343.

Warrington JP， George EM， Palei AC， et al.Recent advances in the understanding of the pathophysiology of preeclampsia［J］.Hypertension， 2013， 62（4）：666-673. doi：10.1161/HYPERTENSIONAHA.113.00588http://dx.doi.org/10.1161/HYPERTENSIONAHA.113.00588.

Betoni JS，Derr K，Pahl MC，et al.MicroRNA analysis in placentas from patients with preeclampsia： comparison of new and published results［J］.Hypertens Pregnancy，2013，32（4）：321-339.doi：10.3109/10641955.2013.807819http://dx.doi.org/10.3109/10641955.2013.807819.

岳巾晶，曾莹，郭晓珮，等.miR-155通过调控PKG1影响滋养细胞生物学功能并参与子痫前期的机制探讨［J］.天津医药，2023，51（9）：928-934.doi：10.11958/20221869http://dx.doi.org/10.11958/20221869.

霍春霞，谢玲，赵得雄.miR-409-3p靶向CXCL9调控滋养层细胞的增殖、迁移和侵袭［J］.免疫学杂志，2021，37（3）：217-223.doi：10.13431/j.cnki.immunol.j.20210032http://dx.doi.org/10.13431/j.cnki.immunol.j.20210032.

Mo W， Jin J， Wang X， et al. MicroRNA-206 contributes to the progression of preeclampsia by suppressing the viability and mobility of trophocytes via the inhibition of AGTR1［J］. Physiol Res， 2023， 72（5）：597-606. doi：10.33549/physiolres.935131http://dx.doi.org/10.33549/physiolres.935131.

Gu Y， Song N， Gu WW， et al. Down-regulation of miR-486-5p expression inhibits the proliferation and invasion of human extravillous trophoblasts， and decreased peripheral plasma miR-486-5p level is associated with human early pregnancy loss［J］. Acta Biochim Biophys Sin （Shanghai）， 2023， 55（11）：1847-1850. doi：10.3724/abbs.2023239http://dx.doi.org/10.3724/abbs.2023239.

Lu TX， Rothenberg ME. MicroRNA［J］. J Allergy Clin Immunol， 2018， 141（4）：1202-1207.doi： 10.1016/j.jaci.2017.08.034http://dx.doi.org/10.1016/j.jaci.2017.08.034.

Ko S， Kim M， Molina L， et al. YAP1 activation and Hippo pathway signaling in the pathogenesis and treatment of intrahepatic cholangiocarcinoma［J］. Adv Cancer Res， 2022，156（1）：283-317. doi： 10.1016/bs.acr.2022.02.003http://dx.doi.org/10.1016/bs.acr.2022.02.003.

Liu R， Wei C， Ma Q， et al. Hippo-YAP1 signaling pathway and severe preeclampsia （sPE） in the Chinese population［J］. Pregnancy Hypertens， 2020， 19（1）：1-10. doi：10.1016/j.preghy.2019.11.002http://dx.doi.org/10.1016/j.preghy.2019.11.002.

Xu B， Geng X， Liu X， et al. Long non-coding RNA FAM99A modulated YAP1 to affect trophoblast cell behaviors in preeclampsia by sponging miR-134-5p［J］. Braz J Med Biol Res， 2020， 53（12）：e9732. doi：10.1590/1414-431X20209732http://dx.doi.org/10.1590/1414-431X20209732.

焦娜，贾源君，范丽丽.重度子痫前期胎盘组织中YAP1、TAZ表达与围生结局的相关性［J］.中国计划生育学杂志，2022，30（11）：2480-2484.doi：10.3969/j.issn.1004-8189.2022.11.012http://dx.doi.org/10.3969/j.issn.1004-8189.2022.11.012.

Hu Y， Ma Y， Luo G， et al. Effect of miR-375 regulates YAP1 on the invasion，apoptosis，and epithelial-mesenchymal transition of cervical cancer HeLa cells［J］. Evid Based Complement Alternat Med， 2021， 2021（1）：3088723. doi：10.1155/2021/3088723http://dx.doi.org/10.1155/2021/3088723.

Wang X， Ou H， Zhou L， et al. Long non-coding RNA LUCAT1 promotes the progression of clear cell renal cell carcinoma via the microRNA-375/YAP1 axis［J］. Exp Ther Med， 2021， 22（1）：754. doi：10.3892/etm.2021.10186http://dx.doi.org/10.3892/etm.2021.10186.

潘怡霞，刘睿，卫婵，等.缺氧环境下YAP1的表达对子痫前期胎盘血管形成的影响及作用机制［J］.西安交通大学学报（医学版），2021，42（4）：547-553.doi：10.7652/jdyxb202104010http://dx.doi.org/10.7652/jdyxb202104010.

Views

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Effect of circDNMT1 regulating miR-377-3p/PUM1 axis on proliferation,apoptosis and epithelial-mesenchymal transition of triple-negative breast cancer cells

lncRNA NEAT1 promoting the proliferation and apoptosis escape of laryngeal papilloma cells by activating BZW2 through the inhibition of miR-let-7a

Mechanism of lncRNA MIR4435-2HG targeting miR-376a-3p to regulate the biological behavior of cholangiocarcinoma cells

Effects and molecular mechanisms of Epac1 on pituitary adenoma cell proliferation and cell cycle

Expression and biological role of DCST1-AS1 in non-small cell lung cancer

Related Author

LIN Yao-ting

LU Hong-quan

ZHENG Mei-ling

LIN Ming-li

ZHANG Bo

WANG Song

Ayiheng· Qukuerhan

YU Zi-dan

Related Institution

Department of General Surgery，Danzhou People's Hospital

Department of Medical Oncology，Hainan Western Central Hospital

Department of Laboratory， Danzhou People's Hospital

Department of Otolaryngology,First Affiliated Hospital of Xinjiang Medical University

Rehabilitation Center， Huizhou Third People's Hospital

⁰