Department of Gynecology and Obstetrics, Münster University Hospital, Münster, Germany.
Fertil Steril. 2013 Apr;99(5):1346-1355.e5. doi: 10.1016/j.fertnstert.2012.11.055. Epub 2013 Jan 8.
To study the function of miR-145, known to be dysregulated in endometriosis, and to identify its target genes in an in vitro endometriosis model.
Experimental laboratory study.
University medical centers.
PATIENT(S): Primary endometrial stroma cells were derived from eutopic endometrium of three American Society for Reproductive Medicine stage III endometriosis patients and from ectopic lesions of four patients with deep infiltrating endometriosis.
INTERVENTION(S): The human endometriotic cell line 12Z and primary eutopic and ectopic endometrial stroma cells were transiently transfected with miR-145 precursors or anti-miR-145 inhibitors and investigated for posttranscriptional regulation of predicted target genes and changes in cell behavior.
MAIN OUTCOME MEASURE(S): Predicted target expression was measured by quantitative reverse transcription-polymerase chain reaction and Western blotting, and altered cell behavior was monitored by cell proliferation assays. The 12Z cells were additionally investigated by Matrigel invasion assays, cell cycle analysis, side population analysis, and aldehyde dehydrogenase activity assays.
RESULT(S): In all cells investigated, miR-145 overexpression inhibited cell proliferation and induced down-regulation of FASCIN-1, SOX2, and MSI2. In 12Z cells miR-145 upregulation increased Matrigel invasiveness and reduced side population and aldehyde dehydrogenase-1 activity. Additional down-regulated targets in 12Z cells included OCT4, KLF4, PODXL, JAM-A, and SERPINE1/PAI-1. ACTG2 and TAGLN were up-regulated upon pre-miR-145 transfection. JAM-A, FASCIN-1, and PAI-I down-regulation in 12Z cells were confirmed by Western blotting.
CONCLUSION(S): miR-145 inhibits endometriotic cell proliferation, invasiveness, and stemness by targeting multiple pluripotency factors, cytoskeletal elements, and protease inhibitors.
研究 miR-145 的功能，已知其在子宫内膜异位症中失调，并在体外子宫内膜异位症模型中鉴定其靶基因。
实验性实验室研究。
大学医学中心。
从三位美国生殖医学协会（ASRM）III 期子宫内膜异位症患者的在位子宫内膜和四位深部浸润性子宫内膜异位症患者的异位病变中获得原代子宫内膜基质细胞。
瞬时转染 miR-145 前体或抗 miR-145 抑制剂，研究预测靶基因的转录后调控和细胞行为的变化。
通过定量逆转录-聚合酶链反应和 Western blot 测量预测靶基因的表达，并通过细胞增殖测定监测改变的细胞行为。还通过 Matrigel 侵袭测定、细胞周期分析、侧群分析和醛脱氢酶活性测定对 12Z 细胞进行了研究。
在所研究的所有细胞中，miR-145 过表达抑制细胞增殖并诱导 FASCIN-1、SOX2 和 MSI2 下调。在 12Z 细胞中，miR-145 的上调增加了 Matrigel 的侵袭能力，并降低了侧群和醛脱氢酶-1 的活性。在 12Z 细胞中，其他下调的靶基因包括 OCT4、KLF4、PODXL、JAM-A 和 SERPINE1/PAI-1。转染前 miR-145 后，ACTG2 和 TAGLN 上调。Western blot 证实了 12Z 细胞中 JAM-A、FASCIN-1 和 PAI-1 的下调。
miR-145 通过靶向多个多能性因子、细胞骨架元件和蛋白酶抑制剂来抑制子宫内膜异位症细胞的增殖、侵袭和干性。
