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. 2019 Apr 20;39(4):437–442. [Article in Chinese] doi: 10.12122/j.issn.1673-4254.2019.04.09

维生素D通过下调miR-21的表达促进人胎盘滋养细胞的迁移和侵袭

Vitamin D down-regulates microRNA-21 expression to promote human placental trophoblast cell migration and invasion in vitro

Zhiyi ZHOU 1, Xiaojuan LI 1, Guoqing JIANG 1, Jue WANG 2,3,4, Yuan QIAN 2,3,4,*
PMCID: PMC6744004  PMID: 31068287

Abstract

Objective

To investigate the effect of vitamin D on microRNA-21(miR-21) expression and migration and invasion of human placental trophoblast cells.

Methods

The changes in the expression of miR-21 were detected using RT-qPCR in HTR-8/SVneo cells following stimulation by vitamin D at different doses for 24, 48 and 72 h.HTR-8/SVneo cells transfected with miR-21 mimic or inhibitor with or without vitamin D treatment were examined for changes in cell migration and invasion abilities using Transwell assay, and Western blotting was used to detect protein expressions of E-cadherin, fibronectin, and MMP9.

Results

Vitamin D obviously inhibited the expression of micoRNA-21 in HTR-8/SVneo cells in a concentration-and time-dependent manner.Transfection with the miR-21 mimic significantly inhibited the migration and invasion of HTR-8/SVneo cells, and this inhibitory effect was abolished by treatment with vitamin D; transfection with miR-21 inhibitor obviously promoted the migration and invasion of HTR-8/SVneo cells, and these effects were not significantly affected by vitamin D treatment.

Conclusion

Vitamin D may promote trophoblast cell migration and invasion to accelerate the development of preeclampsia by down-regulating the expression of miR-21.

Keywords: vitaminD, microRNA-21, trophoblasts, HTR-8/SVneo cells, migration, human placental trophoblast cells

子痫前期（PE）是妊娠期特有疾病，约影响全球5~ 8%的妊娠妇女，是引起孕妇和围产儿死亡的主要原因之一，除了娩出胎盘，目前尚无有效治疗手段[1]。PE发病机制未完全明确，普遍考虑为多因素共同作用的结果，如子宫螺旋动脉重铸障碍，胎盘源性生理物质分泌异常，母体全身炎症反应等[2-4]。

维生素D是人体内不可或缺的类固醇激素，它非经典作用方面的功能越来越受重视，对其抗癌作用以及免疫功能调节分子机制的研究已成为热点[5-6]。越来越多的证据表明维生素D缺乏可能与PE的发病有关[7-8]。MircoRNA-21（miR-21）是目前发现的在恶性肿瘤、心血管疾病、免疫性疾病等多种疾病中均高表达的microRNA（miRNA），被视作一种原癌基因[9-14]。许多报道表明PE患者存在miR-21表达的失调，不过，miR-21在PE患者体内是高表达还是低表达始终未能得到一致的结论[15-19]。研究表明维生素D可通过直接机制（维生素D反应元件VDRE）和间接机制（影响成熟miRNA发生）调节miRNA的表达[20]，通过调节miRNA表达，维生素D参与了多种疾病的发生发展过程[21-24]。已有研究报道指出维生素D可促进绒毛外滋养细胞侵袭[25]，但是，维生素D是否通过靶向miRNA影响PE细胞运动性及其具体机制尚未明确。本研究主要检测了胎盘滋养细胞系中，维生素D水平对miR-21表达情况的影响以及维生素D协同miR-21对滋养细胞迁移侵袭能力的影响，以期进一步探讨二者在PE发病机制中的作用并为PE治疗提供新思路。

1. 材料和方法

1.1. 细胞株及主要试剂

人胎盘滋养细胞株HTR-8/SVneo（HTR）购自BioVector质粒载体菌种细胞基因保藏中心，RPMI 1640培养基、Opti-MEM培养液（Gibco），胎牛血清（BD）。1, 25二羟维生素D3[1, 25(OH)2D3]（Sigma），miR-21 mimic、miR-21 inhibitor及其对应的miRNA阴性对照均由上海吉玛制药技术有限公司合成。转染试剂Lipofectamine2000（Invitrogen），RNA提取逆转录和实时荧光定量所用试剂（QIAGEN），miR-21引物和内参U6引物（吉码）。Transwell小室和BD基质胶（Corning），小鼠抗人GAPDH抗体、兔抗人MMP9抗体（Santa Cruz），兔抗人E-cadherin、Fibronectin抗体（Abcam）。

1.2. 方法

1.2.1. 细胞培养以及1, 25(OH)2D3的刺激和转染

HTR细胞在含10%胎牛血清的RPMI 1640培养基中，37 ℃、5% CO2无菌培养箱中培养，每2~3 d传代1次，取对数期生长细胞进行实验。细胞融合度达80%左右后按转染试剂说明书进行转染；若需联合1, 25(OH)2D3刺激细胞，则在转染6h换培养基时加1, 25(OH)2D3。

1.2.2. RT-qPCR法检测miR-21表达情况

1, 25(OH)2D3以无水乙醇配制成100 µmol/L的贮存液，-20 ℃避光保存。取对数期生长细胞，胰酶消化后制备成单细胞悬液，接种到6孔板，密度为2×105/孔。待细胞贴壁后，实验组加1, 25(OH)2D3, 工作浓度分别为0.1、1、10 nmol/L，并设空白对照组，培养箱中分别培养24、48、72 h，按试剂盒说明书提RNA，RT-qPCR实验检测miR-21表达情况。以U6作为参考，所用引物序列如下：

miR-21：F primer：ACGTTGTGTAGCTTATCAGACTG

R primer：AATGGTTGTTCTCCACACTCTC

U6：F primer：CAGCACATATACTAAAATTGGAACG

R primer：ACGAATTTGCGTGTCATCC

1.2.3. Transwell检测细胞迁移和侵袭能力

取24孔板Transwell小室，上室中加入单独转染miR-21 mimic/ inhibitor或联合10 nmol的1, 25(OH)2D3处理的HTR细胞悬液（浓度为2.5×105/mL）200 μL，下室中加入含10%胎牛血清的RPMI 1640培养基600 μL，37 ℃培养箱继续培养24 h后，用棉签擦拭掉上室中未穿过膜的细胞，PBS冲洗小室3次，甲醇固定约20 min，0.1%结晶紫溶液中染色10 min后，倒置显微镜下随机选取5个视野，200倍光学显微镜下拍照计数每个视野内穿透小室微孔膜的细胞数，计算平均数，随后做统计学分析。细胞侵袭实验的Transwell小室上室提前加入100 μL按说明书稀释后的Matrigel胶，细胞接种数目为1×105/孔，培养时间为36~48 h，其余操作同迁移实验。

1.2.4. Western blotting检测EMT相关蛋白的表达水平

取对数生长期细胞，经相应处理后，提取蛋白质，通过SDS-PAGE凝胶电泳分离，将蛋白转移到硝酸纤维膜上，4 ℃封闭过夜后加入一抗，4 ℃过夜后加入二抗，室温下孵育3 h。用化学发光试剂增强反应，以凝胶成像系统分析结果。

1.3. 统计学处理

采用GraphPad Prism7软件进行统计学分析，对两组数据比较采用t检验，实验数据以均数±标准差表示；对多组数据比较采用单因素方差分析，实验数据以95% CI表示。P < 0.05为差异具有统计学意义。

2. 结果

2.1. 1, 25(OH)2D3可抑制HTR细胞miR-21的表达

不同浓度1, 25(OH)2D3作用细胞株后抑制了miR-21的表达，抑制作用呈浓度-时间依赖性增强（图 1）。

1.

1

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不同浓度1, 25(OH)2D3刺激HTR细胞24、48、72 h后miR-21的表达水平

Expression levels of miR-21 in HTR cells after stimulation by 0, 0.1, 1 and 10 nmol/L 1, 25(OH)2D3 for 24, 48 and 72 h (*P < 0.05; **P < 0.01 vs control).

2.2. miR-21过表达抑制HTR细胞迁移和侵袭

与阴性对照相比，转染miR-21 mimic后HTR细胞的迁移和侵袭能力受到抑制（P < 0.05）。与阴性对照组相比，转染miR-21 inhibitor后促进了HTR细胞的迁移和侵袭能力（P < 0.05，图 2）。

2.

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Transwell实验检测miR-21 mimic /inhibitor刺激HTR细胞后对HTR的迁移、侵袭能力的影响及统计分析

Transwell assayfor assessing the migration and invasion of HTR cells after transefection with the miR-21 mimic and inhibitor (A, C) and quantitative analysis of the results (B, D). *P < 0.05.

2.3. 1, 25(OH)2D3可减弱miR-21过表达对HTR细胞迁移、侵袭的抑制作用

与单独转染miR-21mimic组相比，miR-21mimic联合1, 25(OH)2D3刺激细胞减弱了miR-21过表达对HTR细胞迁移和侵袭的抑制作用（P < 0.05）。与inhibitor组相比，miR-21 inhibitor联合1, 25(OH)2D3刺激则与单独转染miR-21 inhibitor对HTR细胞的迁移、侵袭能力无明显改变（P>0.05）。

3.

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Transwell实验检测miR-21 mimic/inhibitor联合1, 25(OH)2D3刺激HTR细胞后对HTR的迁移、侵袭能力的影响及统计分析

Transwell assayfor assessing the migration and invasion of HTR cells after transefection with the miR-21 mimic and inhibitor in the presence or absence of 1, 25(OH)2D3 (A, C) and quantitative analysis of the results (B, D) (*P < 0.05, **P < 0.01). Transwell assayfor assessing the migration and invasion of HTR cells after transefection with the miR-21 mimic and inhibitor (A, C) and quantitative analysis of the results (B, D). *P < 0.05.

2.4. miR-21上调E-cadherin，下调fibronectin、MMP9蛋白的表达

WB结果显示，HTR细胞转染miR-21mimic后，上皮标记物E-cadherin蛋白表达水平增加，间充质标记物fibronectin蛋白表达水平降低，MMP9表达水平降低。瞬时转染miR-21 inhibitor则结果相反（图 4）。

4.

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Western blot检测miR-21mimic/inhibitor刺激HTR后EMT相关蛋白的表达情况

Western blot analysis of expressions of EMT-related proteins in HTR cell after transfection with the miR-21 mimic or inhibitor (*P < 0.05, **P < 0.01).

2.5. 1, 25(OH)2D3可减弱miR-21对E-cadherin、fibronectin、MMP9调控作用

按要求对细胞经相应处理后，WB结果显示，联合1, 25(OH)2D3后，与单独转染miR-21mimic相比，Ecadherin蛋白表达降低，Fibronectin、MMP9蛋白表达增加；与单独转染miR-21inhibitor相比，蛋白表达水平无明显变化（图 5）。

5.

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Western blot检测miR-21mimic/inhibitor联合维生素D刺激HTR后EMT相关蛋白的表达情况

Western blot analysis of the expressions of EMT-related proteins in HTR cells after transfection with the miR-21 mimic/inhibitor in the presnce or absence of vitamin D (*P < 0.05).

3. 讨论

近年来关于维生素D对人类机体健康影响的研究有了明显的进步。除了经典的调节钙磷代谢、影响骨骼发育外，它的抗炎症和抗癌特性越发引起人们的关注。目前许多miRNA被鉴定为维生素D靶标，如miR-17/ 22/100/146/155/21等[21, 24-25]。虽然，关于维生素D调控miRNA表达的研究较多，但有关维生素D在PE疾病中对miR-21表达的调控以及二者在PE发病机制中起何种作用的研究还不得而知。

在前期研究中，我们发现与健康妊娠妇女相比，PE患者存在维生素D水平降低和胎盘miR-21表达水平升高的现象。本次实验中，我们使用最常见的维生素D活化型产物—1, 25(OH)2D3刺激HTR细胞，发现1, 25(OH)2D3可抑制HTR细胞miR-21的表达，且抑制效果呈一定的浓度-时间依赖性增强。这表明1, 25(OH)2D3可能通过减弱PE患者高表达的miR-21从而影响PE的病理过程。

大量研究表明miR-21可通过不同机制调节多种癌细胞的侵袭性[10-12, 26-27]，考虑到胎盘滋养细胞侵袭能力不足是PE发病的重要原因之一，我们推测胎盘中异常表达的miR-21也可能参与到滋养细胞的运动过程。于是，我们进行transwell实验研究了miR-21单独或联合1, 25(OH)2D3共同刺激细胞时对细胞迁移和侵袭的影响。结果显示，与NC组相比，转染miR-21mimic后抑制了HTR细胞的迁移和侵袭能力，当联合1, 25(OH)2D3后减弱了该抑制作用。转染miR-21 inhibitor后可促进HTR细胞的迁移与侵袭能力，然而，用1, 25(OH)2D3联合miR-21 inhibitor共同刺激细胞时，与单独miR-21 inhibitor刺激相比，细胞迁移和侵袭能力无明显变化。不同的刺激因素可能造成相似的细胞功能表现，但它们作用于细胞时具体激活了细胞的哪条通路以及miR-21抑制HTR细胞运动能力的具体机制，我们还不得而知。此外，1, 25(OH)2D3与miR-21 inhibitor两种处理方式均可抑制miR-21水平，细胞受外界刺激引起相应改变是要在一定的阈值以内的，当miRNA水平被一种刺激因素抑制到一个临界值时，继续增加刺激或许并不会有太明显的变化。这也许是此次研究中未能出现预期的1, 25(OH)2D3联合miR-21 inhibitor可协同促进HTR细胞迁移和侵袭的原因。

E-cadherin在母-胎界面的蜕膜上皮细胞及滋养细胞广泛表达，赵洪波等人研究表明高表达的E-cadherin可抑制滋养细胞的侵袭性[28]。Fibronectin在细胞粘附、生长和迁移中发挥重要作用，其表达水平与PE的严重程度具有一定的正相关性[29-30]。MMP9主要由滋养细胞分泌，它的表达有助于滋养细胞的侵袭和细胞外基质的溶解，MMP9异常可影响滋养细胞的侵袭能力，导致血管重铸不全[4, 31]。因此我们WB验证了细胞经相应处理后这些蛋白的表达情况。实验结果显示，miR-21过表达能上调E-cadherin，下调Fibronectin、MMP9蛋白的表达，而1, 25(OH)2D3能减弱该调控作用，转染miR-21 inhibitor则出现相反的结果，这与transwell实验结果相对应。进一步阐明了1, 25(OH)2D3可能通过抑制miR-21表达，引起EMT相关蛋白和MMP9蛋白表达的改变，进而影响滋养细胞的运动能力。

本次研究也存在一些不足之处。滋养细胞有JEG、HTR、BeWo、人绒毛膜滋养细胞4种细胞系，此次实验目前只选择了HTR这一种细胞株，尚需同步其他几种滋养细胞株实验进行验证。此外，我们仅在细胞水平进行了验证，补充维生素D是否能通过降低miR-21来缓解PE的临床症状或疾病进展，以到达治疗PE的目的，这还需要进一步的动物实验加以验证。

综上所述，维生素D和miR-21与PE的发生、发展关系密切。维生素D可能通过抑制miR-21的表达促进滋养细胞的迁移与侵袭，从而影响PE的病理过程。本实验为维生素D对PE作用的进一步研究及应用于临床提供实验依据。随着研究的深入，维生素D在PE的临床防治上将会有更广泛的作用。

Biography

周芝熠, 硕士, E-mail: 164930412@qq.com

Funding Statement

国家自然科学基金（81360103，81760273）；云南省高层次卫生技术人才培养（后备人才）（H201628）；云南省教育厅科学研究基金项目（2016ZZX090）；2017年云南省应用基础研究计划面上项目（2017FB107）

Supported by National Natural Science Foundation of China (81360103, 81760273)

Contributor Information

周 芝熠 (Zhiyi ZHOU), Email: 164930412@qq.com.

钱 源 (Yuan QIAN), Email: 549978781@qq.com.

References

  • 1.Ahmed A, Rezai H, Broadway-Stringer S. Evidence-Based Revised View of the Pathophysiology of Preeclampsia. https://www.ncbi.nlm.nih.gov/pubmed/27873232. Adv Exp Med Biol. 2017;956:355–74. doi: 10.1007/5584_2016_168. [Ahmed A, Rezai H, Broadway-Stringer S.Evidence-Based Revised View of the Pathophysiology of Preeclampsia[J].Adv Exp Med Biol, 2017, 956:355-74.] [DOI] [PubMed] [Google Scholar]
  • 2.Falco ML, Sivanathan J, Laoreti A, et al. Placental histopathology associated with pre-eclampsia:systematic review and meta-analysis. Ultrasound Obstet Gynecol. 2017;50(3):295–301. doi: 10.1002/uog.17494. [Falco ML, Sivanathan J, Laoreti A, et al.Placental histopathology associated with pre-eclampsia:systematic review and meta-analysis[J].Ultrasound Obstet Gynecol, 2017, 50(3):295-301.] [DOI] [PubMed] [Google Scholar]
  • 3.伍 天文, 王 钢, 李 晓青, et al. 子痫前期发病机制的研究现状. http://d.old.wanfangdata.com.cn/Periodical/hxykdxxb201501021. 基因组学与应用生物学. 2017;36(3):1266–71. [伍天文, 王钢, 李晓青, 等.子痫前期发病机制的研究现状[J].基因组学与应用生物学, 2017, 36(3):1266-71.] [Google Scholar]
  • 4.孙 雯, 陈 敦金. 胎盘功能与子痫前期的研究进展. 中华产科急救电子杂志. 2013;2(1):51–4. doi: 10.3877/cma.j.issn.2095-3259.2013.01.015. [孙雯, 陈敦金.胎盘功能与子痫前期的研究进展[J].中华产科急救电子杂志, 2013, 2(1):51-4.] [DOI] [Google Scholar]
  • 5.Zeljic K, Supic G, Magic Z. New insights into vitamin D anticancer properties:focus on miRNA modulation. Mol Genet Genomics. 2017;292(3):511–24. doi: 10.1007/s00438-017-1301-9. [Zeljic K, Supic G, Magic Z.New insights into vitamin D anticancer properties:focus on miRNA modulation[J].Mol Genet Genomics, 2017, 292(3):511-24.] [DOI] [PubMed] [Google Scholar]
  • 6.Chen DJ, Li LJ, Yang XK, et al. Altered microRNAs expression in T cells of patients with SLE involved in the lack of vitamin D. https://www.ncbi.nlm.nih.gov/pubmed/28977929. Oncotarget. 2017;8(37):62099–110. doi: 10.18632/oncotarget.19062. [Chen DJ, Li LJ, Yang XK, et al.Altered microRNAs expression in T cells of patients with SLE involved in the lack of vitamin D[J].Oncotarget, 2017, 8(37):62099-110.] [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 7.蒲 元芳, 谢 艳, 战 玉峰, et al. 孕妇血清25羟基维生素D水平与妊娠合并症的关系. 生殖医学杂志. 2015;24(1):79–83. doi: 10.3969/j.issn.1004-3845.2015.01.018. [蒲元芳, 谢艳, 战玉峰, 等.孕妇血清25羟基维生素D水平与妊娠合并症的关系[J].生殖医学杂志, 2015, 24(1):79-83.] [DOI] [Google Scholar]
  • 8.邓 娜, 范 翠芳. 维生素D对子痫前期的影响及作用机制的研究进展. 安徽医药. 2018;22(8):1441–4. doi: 10.3969/j.issn.1009-6469.2018.08.003. [邓娜, 范翠芳.维生素D对子痫前期的影响及作用机制的研究进展[J].安徽医药, 2018, 22(8):1441-4.] [DOI] [Google Scholar]
  • 9.Feng YH, Tsao CJ. Emerging role of microRNA-21 in cancer. Biomed Rep. 2016;5(4):395–402. doi: 10.3892/br.2016.747. [Feng YH, Tsao CJ.Emerging role of microRNA-21 in cancer[J].Biomed Rep, 2016, 5(4):395-402.] [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 10.Fang H, Xie JP, Zhang M, et al. miRNA-21 promotes proliferation and invasion of triple-negative breast cancer cells through targeting PTEN. https://www.ncbi.nlm.nih.gov/pubmed/28386324. Am J Transl Res. 2017;9(3):953–61. [Fang H, Xie JP, Zhang M, et al.miRNA-21 promotes proliferation and invasion of triple-negative breast cancer cells through targeting PTEN[J].Am J Transl Res, 2017, 9(3):953-61.] [PMC free article] [PubMed] [Google Scholar]
  • 11.Cao J, Liu J, Xu R, et al. MicroRNA-21 stimulates epithelial-tomesenchymal transition and tumorigenesis in clear cell renal cells. Mol Med Rep. 2016;13(1):75–82. doi: 10.3892/mmr.2015.4568. [Cao J, Liu J, Xu R, et al.MicroRNA-21 stimulates epithelial-tomesenchymal transition and tumorigenesis in clear cell renal cells[J].Mol Med Rep, 2016, 13(1):75-82.] [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 12.Wu Y, Song Y, Xiong Y, et al. MicroRNA-21(Mir-21) promotes cell growth and invasion by repressing tumor suppressor PTEN in colorectal cancer. Cell Physiol Biochem. 2017;43(3):945–58. doi: 10.1159/000481648. [Wu Y, Song Y, Xiong Y, et al.MicroRNA-21(Mir-21) promotes cell growth and invasion by repressing tumor suppressor PTEN in colorectal cancer[J].Cell Physiol Biochem, 2017, 43(3):945-58.] [DOI] [PubMed] [Google Scholar]
  • 13.Sheane BJ, Smyth P, Scott K, et al. An association between MicroRNA-21 expression and vitamin D deficiency in coronary artery disease. Microrna. 2015;4(1):57–63. doi: 10.2174/2211536604666150414203919. [Sheane BJ, Smyth P, Scott K, et al.An association between MicroRNA-21 expression and vitamin D deficiency in coronary artery disease[J].Microrna, 2015, 4(1):57-63.] [DOI] [PubMed] [Google Scholar]
  • 14.Liu P, Wheelwright M, Teles R, et al. MicroRNA-21 targets the vitamin D-dependent antimicrobial pathway in leprosy. https://www.ncbi.nlm.nih.gov/pubmed/22286305. J Immunol. 2011;186(1):267–73. doi: 10.1038/nm.2584. [Liu P, Wheelwright M, Teles R, et al.MicroRNA-21 targets the vitamin D-dependent antimicrobial pathway in leprosy[J].J Immunol, 2011, 186(1):267-73.] [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 15.Jairajpuri DS, Malalla ZH, Mahmood NA. Circulating microRNA expression as predictor of preeclampsia and its severity. Gene. 2017;627:543–8. doi: 10.1016/j.gene.2017.07.010. [Jairajpuri DS, Malalla ZH, Mahmood NA.Circulating microRNA expression as predictor of preeclampsia and its severity[J].Gene, 2017, 627:543-8.] [DOI] [PubMed] [Google Scholar]
  • 16.Svecova I, Vazan M, Zubor P, et al. Overexpression of miR-21 and miR-122 in preeclamptic placentas. https://www.ncbi.nlm.nih.gov/pubmed/26859593. Neuro Endocrinol Lett. 2015;36(7):695–9. [Svecova I, Vazan M, Zubor P, et al.Overexpression of miR-21 and miR-122 in preeclamptic placentas[J].Neuro Endocrinol Lett, 2015, 36(7):695-9.] [PubMed] [Google Scholar]
  • 17.曾 辛, 王 枫, 黄 齐香, et al. 重度子痫前期患者胎盘组织中miR-21的表达及其临床意义. http://d.old.wanfangdata.com.cn/Periodical/zgfybj201214042. 中国妇幼保健. 2012;27(14):2184–6. [曾辛, 王枫, 黄齐香, 等.重度子痫前期患者胎盘组织中miR-21的表达及其临床意义[J].中国妇幼保健, 2012, 27(14):2184-6.] [Google Scholar]
  • 18.Choi SY, Yun J, Lee OJ, et al. MicroRNA expression profiles in placenta with severe preeclampsia using a PNA-based microarray. Placenta. 2013;34(9):799–804. doi: 10.1016/j.placenta.2013.06.006. [Choi SY, Yun J, Lee OJ, et al.MicroRNA expression profiles in placenta with severe preeclampsia using a PNA-based microarray[J].Placenta, 2013, 34(9):799-804.] [DOI] [PubMed] [Google Scholar]
  • 19.Pillar N, Yoffe L, Hod M, et al. The possible involvement of microRNAs in preeclampsia and gestational diabetes mellitus. Best Pract Res Clin Obstet Gynaecol. 2015;29(2):176–82. doi: 10.1016/j.bpobgyn.2014.04.021. [Pillar N, Yoffe L, Hod M, et al.The possible involvement of microRNAs in preeclampsia and gestational diabetes mellitus[J].Best Pract Res Clin Obstet Gynaecol, 2015, 29(2):176-82.] [DOI] [PubMed] [Google Scholar]
  • 20.Ma YY, Trump DL, Johnson CS. Vitamin D and miRNAs in cancer. Curr Gene Ther. 2014;14(4):269–75. doi: 10.2174/1566523214666140612153537. [Ma YY, Trump DL, Johnson CS.Vitamin D and miRNAs in cancer[J].Curr Gene Ther, 2014, 14(4):269-75.] [DOI] [PubMed] [Google Scholar]
  • 21.Karkeni E, Bonnet L, Marcotorchino J, et al. Vitamin D limits inflammation-linked microRNA expression in adipocytes in vitro and in vivo:A new mechanism for the regulation of inflammation by vitamin D. Epigenetics. 2018;13(2):156–62. doi: 10.1080/15592294.2016.1276681. [Karkeni E, Bonnet L, Marcotorchino J, et al.Vitamin D limits inflammation-linked microRNA expression in adipocytes in vitro and in vivo:A new mechanism for the regulation of inflammation by vitamin D[J].Epigenetics, 2018, 13(2):156-62.] [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 22.Duan X, Guan Y, Li Y, et al. Vitamin D Potentiates the Inhibitory Effect of MicroRNA-130a in Hepatitis C Virus Replication Independent of Type Ⅰ Interferon Signaling Pathway. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4427768/ Mediators Inflamm. 2015;2015:508989. doi: 10.1155/2015/508989. [Duan X, Guan Y, Li Y, et al.Vitamin D Potentiates the Inhibitory Effect of MicroRNA-130a in Hepatitis C Virus Replication Independent of Type Ⅰ Interferon Signaling Pathway[J].Mediators Inflamm, 2015, 2015:508989.] [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 23.Alvarez-Diaz S, Valle N, Ferrer-Mayorga GA, et al. MicroRNA-22 is induced by vitamin D and contributes to its antiproliferative, antimigratory and gene regulatory effects in colon cancer cells. Hum Mol Genet. 2012;21(10):2157–65. doi: 10.1093/hmg/dds031. [Alvarez-Diaz S, Valle N, Ferrer-Mayorga GA, et al.MicroRNA-22 is induced by vitamin D and contributes to its antiproliferative, antimigratory and gene regulatory effects in colon cancer cells[J].Hum Mol Genet, 2012, 21(10):2157-65.] [DOI] [PubMed] [Google Scholar]
  • 24.Yu H, Xu M, Dong YZ, et al. 1, 25(OH)(2) D-3 attenuates pulmonary arterial hypertension via microRNA-204 mediated Tgfbr2/Smad signaling. Exp Cell Res. 2018;362(2):311–23. doi: 10.1016/j.yexcr.2017.11.032. [Yu H, Xu M, Dong YZ, et al.1, 25(OH)(2) D-3 attenuates pulmonary arterial hypertension via microRNA-204 mediated Tgfbr2/Smad signaling[J].Exp Cell Res, 2018, 362(2):311-23.] [DOI] [PubMed] [Google Scholar]
  • 25.Ma Y, Hu Q, Luo W, et al. 1α, 25(OH)2D3 differentially regulates miRNA expression in human bladder cancer cells. J Steroid Biochem Mol Biol. 2015;148:166–71. doi: 10.1016/j.jsbmb.2014.09.020. [Ma Y, Hu Q, Luo W, et al.1α, 25(OH)2D3 differentially regulates miRNA expression in human bladder cancer cells[J].J Steroid Biochem Mol Biol, 2015, 148:166-71.] [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 26.Su CY, Cheng X, Li YS, et al. MiR-21 improves invasion and migration of drug-resistant lung adenocarcinoma cancer cell and transformation of EMT through targeting HBP1. Cancer Med. 2018;7(6):2485–503. doi: 10.1002/cam4.2018.7.issue-6. [Su CY, Cheng X, Li YS, et al.MiR-21 improves invasion and migration of drug-resistant lung adenocarcinoma cancer cell and transformation of EMT through targeting HBP1[J].Cancer Med, 2018, 7(6):2485-503.] [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 27.Luo G, Luo W, Sun X, et al. MicroRNA21 promotes migration and invasion of glioma cells via activation of Sox2 and betacatenin signaling. Mol Med Rep. 2017;15(1):187–93. doi: 10.3892/mmr.2016.5971. [Luo G, Luo W, Sun X, et al.MicroRNA21 promotes migration and invasion of glioma cells via activation of Sox2 and betacatenin signaling[J].Mol Med Rep, 2017, 15(1):187-93.] [DOI] [PMC free article] [PubMed] [Google Scholar] [Retracted]
  • 28.赵 洪波, 薛 立荣, 李 大金. E-钙黏蛋白(E-cadheri)对人滋养细胞侵袭能力及妊娠结局的影响. http://d.old.wanfangdata.com.cn/Periodical/szyby201201003. 生殖与避孕. 2012;32(1):10–6, 37. [赵洪波, 薛立荣, 李大金.E-钙黏蛋白(E-cadheri)对人滋养细胞侵袭能力及妊娠结局的影响[J].生殖与避孕, 2012, 32(1):10-6, 37.] [Google Scholar]
  • 29.杨 铧琦, 蒋 红. PE患者血清FN及sFlt-1的水平及临床意义研究. http://www.cnki.com.cn/Article/CJFDTotal-YXYZ201606048.htm. 医学研究杂志. 2016;45(6):175–9. [杨铧琦, 蒋红.PE患者血清FN及sFlt-1的水平及临床意义研究[J].医学研究杂志, 2016, 45(6):175-9.] [Google Scholar]
  • 30.Griggs LA, Hassan NT, Malik RS, et al. Fibronectin fibrils regulate TGF-beta 1-induced epithelial-mesenchymal transition. https://www.ncbi.nlm.nih.gov/pubmed/28109697. Matrix Biol. 2017;60(SI):157–75. doi: 10.1016/j.matbio.2017.01.001. [Griggs LA, Hassan NT, Malik RS, et al.Fibronectin fibrils regulate TGF-beta 1-induced epithelial-mesenchymal transition[J].Matrix Biol, 2017, 60-61(SI):157-75.] [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 31.Sakowicz A, Lisowska M, Biesiada LA, et al. Association of maternal and fetal Single-Nucleotide polymorphisms in metalloproteinase (MMP1, MMP2, MMP3, and MMP9) genes with preeclampsia. http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5835279/ Dis Markers. 2018;2018:1371425. doi: 10.1155/2018/1371425. [Sakowicz A, Lisowska M, Biesiada LA, et al.Association of maternal and fetal Single-Nucleotide polymorphisms in metalloproteinase (MMP1, MMP2, MMP3, and MMP9) genes with preeclampsia[J].Dis Markers, 2018, 2018:1371425.] [DOI] [PMC free article] [PubMed] [Google Scholar]

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