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LIM激酶1过表达对子宫内膜异位症来源的基质细胞行为的影响。

Effect of LIM kinase 1 overexpression on behaviour of endometriosis-derived stromal cells.

作者信息

Zhang Zhifang, Chen Peng, Guo Cuishan, Meng Xiannan, Wang Danbo

机构信息

Department of Obstetrics and Gynecology, Shengjing Hospital of China Medical University, 36 Sanhao Street, Shenyang, 110004, People's Republic of China.

出版信息

Cell Tissue Res. 2015 Mar;359(3):885-93. doi: 10.1007/s00441-014-2068-5. Epub 2014 Dec 23.

Abstract

LIM kinase 1 (LIMK1) has been implicated in tumour invasion and migration in several tumour types. However, its role in the progression of endometriosis has not yet been studied. Our aim is to analyse LIMK1 expression in endometriosis-derived stromal cells and to explore the effects of LIMK1 overexpression on their biological behaviour. The mRNA and protein expression levels of LIMK1 of eutopic endometrial stromal cells (ESCs) separated from 30 endometriosis patients and normal endometrial stromal cells (NSCs) separated from 30 patients without endometriosis were analysed by real-time polymerase chain reaction and Western blot. Lentiviral particles containing human LIMK1 short interfering RNA were used to silence the LIMK1 gene in ESCs and LIMK1-expressing lentiviral particles containing the total LIMK1 cDNA was transfected into NSCs to upregulate LIMK1 expression. Cell migration, invasion, proliferation and expression of markers of adhesion, invasion and angiogenesis of ESCs and NSCs were evaluated under basal conditions and after transfection in vitro. The mRNA and protein expression levels of LIMK1 in ESCs were higher than those in NSCs. Under basal conditions, ESCs exhibited greater cell migration, invasion and proliferation and higher levels of markers of adhesion, invasion and angiogenesis than NSCs. The behaviour of ESCs was decreased after LIMK1 gene silencing and that of NSCs was elevated after LIMK1 gene upregulation. Thus, LIMK1 is overexpressed in ESCs thereby facilitating malignant-like behaviour, including enhanced migration, invasion, proliferation and angiogenesis, all of which contribute to the occurrence and development of endometriosis.

摘要

LIM激酶1(LIMK1)已被证明在多种肿瘤类型的侵袭和迁移中发挥作用。然而,其在子宫内膜异位症进展中的作用尚未得到研究。我们的目的是分析LIMK1在子宫内膜异位症来源的基质细胞中的表达,并探讨LIMK1过表达对其生物学行为的影响。通过实时聚合酶链反应和蛋白质免疫印迹法分析了从30例子宫内膜异位症患者分离出的在位子宫内膜基质细胞(ESC)和从30例非子宫内膜异位症患者分离出的正常子宫内膜基质细胞(NSC)中LIMK1的mRNA和蛋白质表达水平。使用含有人类LIMK1短发夹RNA的慢病毒颗粒沉默ESC中的LIMK1基因,并将含有全长LIMK1 cDNA的表达LIMK1的慢病毒颗粒转染到NSC中以上调LIMK1表达。在基础条件下和体外转染后,评估了ESC和NSC的细胞迁移、侵袭、增殖以及黏附、侵袭和血管生成标志物的表达。ESC中LIMK1的mRNA和蛋白质表达水平高于NSC。在基础条件下,ESC比NSC表现出更强的细胞迁移、侵袭和增殖能力,以及更高水平的黏附、侵袭和血管生成标志物。LIMK1基因沉默后ESC的行为减弱,LIMK1基因上调后NSC的行为增强。因此,LIMK1在ESC中过表达,从而促进了类似恶性的行为,包括增强的迁移、侵袭、增殖和血管生成,所有这些都有助于子宫内膜异位症的发生和发展。

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