上海交通大学学报(医学版) ›› 2017, Vol. 37 ›› Issue (10): 1350-.doi: 10.3969/j.issn.1674-8115.2017.10.008

• 论著(基础研究) • 上一篇    下一篇

miR-137过表达滋养细胞对血管内皮细胞生物学功能的调节作用#br#

彭海燕,李华萍   

  1. 上海交通大学附属第六人民医院妇产科,上海 200233
  • 出版日期:2017-10-28 发布日期:2017-11-01
  • 通讯作者: 李华萍,电子信箱:hpli819@126.com
  • 作者简介:彭海燕(1991—),女,硕士生;电子信箱:haiyan5_peng@163.com

Regulation of miR-137 overexpressed trophoblast cells on biological functions of vascular endothelial cells#br#

PENG Hai-yan, LI Hua-ping   

  1. Department of Gynecology and Obstetrics, Shanghai Sixth People’s Hospital, Shanghai Jiao Tong University, Shanghai 200233, China
  • Online:2017-10-28 Published:2017-11-01

摘要:  目的 · 探讨滋养细胞过表达 miR-137 后对血管内皮细胞生物学功能的影响。方法 · 构建 Up-LV-miR-137 和 LV-miR-NC 慢病毒 载体,分别转染滋养细胞 HTR-8/SVneo 后通过 PCR 验证转染效率。成功转染后的滋养细胞与血管内皮细胞共培养,或以转染后滋养 细胞培养基上清液培养血管内皮细胞,CCK8 法、划痕试验和 Transwell 迁移试验分别检测内皮细胞的增殖活性、迁移能力和对单核细 胞 U937 的招募能力。结果 · 成功获得 miR-137 过表达滋养细胞。CCK8 细胞活性检测结果显示,与 miR-137 过表达滋养细胞共培养 或以其培养基上清液培养,血管内皮细胞增殖活性显著降低,高糖环境下增殖活性进一步降低。划痕试验中,miR-137 过表达细胞培 养基使得内皮细胞划痕修复速度减慢。Transwell 迁移试验结果发现,内皮细胞与 miR-137 过表达滋养细胞共培养后,该组单核细胞穿 过小室的细胞数更多,趋化指数更高。结论 · miR-137 过表达滋养细胞可调节血管内皮细胞的生物学功能,降低增殖活性和迁移能力, 增强其对单核细胞的招募能力。

关键词: &ensp, miR-137, 滋养细胞, 血管内皮细胞, 妊娠期糖尿病, 单核细胞

Abstract:

Objective · To study the effect of miR-137 overexpressed trophoblast cells on the biological functions of vascular endothelial cells.  Methods · Lentivirus vectors Up-LV-miR-137 and LV-miR-NC were constructed and transfected into the trophoblast cells HTR-8/SVneo, respectively. And transfection efficiency was verified by PCR. Vascular endothelial cells were cultured with transfected trophoblast cells or their culture medium supernatant. Proliferation, migration activity and ability to recruit monocytes U937 of endothelial cells were detected by CCK8 method, scratch test and Transwell migration test, respectively.  Results · miR-137 overexpressed trophoblast cells were obtained. The results of CCK8 test showed that proliferation of vascular endothelial cells cultured with miR-137 overexpressed trophoblast cells or their culture medium supernatant was inhibited, especially in high glucose condition. The repair of endothelial cells in the transfected cells culture medium slowed down in the scratch test. Transwell migration test results showed that after the endothelial cells were cultured with miR-137 overexpressed trophoblast cells, more monocytes passed through the chambers with higher chemotactic index.  Conclusion · miR-137 overexpressed trophoblast cells can regulate the biological functions of vascular endothelial cells, i.e. reduce proliferation and migration activity and enhance ability to recruit monocytes.

Key words:  miR-137, trophoblast cell, vascular endothelial cell, gestational diabetes mellitus, monocyte