TGF-β1及其抑制剂培养的子宫内膜异位症患者异位子宫内膜间质细胞增殖、侵袭情况观察  

Expression of TGF-β1 mRNA in ectopic endometrium of patients with endometriosis and the proliferation and invasion of endometrial stromal cells cultured with TGF-β1 and its inhibitor

作　　者：于克 王祝荣 YU Ke;WANG Zhurong(The Affiliated Hospital of Yangzhou University, Yangzhou 225000, China)

机构地区：[1]扬州大学附属医院

出　　处：《山东医药》2019年第18期10-14,共5页Shandong Medical Journal

摘　　要：目的观察子宫内膜异位症(Endometriosis, EMs)患者异位子宫内膜组织转化生长因子β 1 (Transforming Growth Factor beta1, TGF-β 1)mRNA的表达变化,观察加入TGF-β 1及其抑制剂培养的子宫内膜间质细胞(endometrial stromal cells, ESCs)增殖、侵袭能力变化,并探讨可能作用机制。方法 100例EMs患者的异位子宫内膜组织及40例未患有EMs患者的正常子宫内膜组织,采用 qRT-PCR法检测TGF-β 1 mRNA。取异位子宫内膜新鲜组织,分离、培养、鉴定原代ESCs;取对数生长期ESCs分为1、2、3、4组,1、2组分别加入终浓度为2 ng/mL的TGF-β 1因子、4 μL PBS(TGF-β 1因子配制溶剂),3、4组分别加入终浓度为1 μmol/L的TGF-β 1抑制剂Ly364947、2 μL DMSO(Ly364947配制溶剂),培养48 h时采用 qRT-PCR法检测各组细胞TGF-β 1 mRNA,分别于培养0、24、48、72 h时采用MTS法测算各组细胞增殖能力(OD值表示),培养48 h时采用Boyden实验观察各组细胞侵袭能力,培养48 h时采用Western blotting法检测各组细胞β-连环蛋白(β-catenin)、cMYC蛋白。结果 EMs患者异位子宫内膜组织、正常子宫内膜组织TGF-β 1 mRNA相对表达量分别为2.26±0.96、1.02±0.38( t =7.908, P <0.05)。培养48 h时1、2组细胞TGF-β 1 mRNA相对表达量分别为8.88±1.03、1.00±0.02( P <0.05),3、4组细胞TGF-β 1 mRNA相对表达量分别为0.57±0.07、1.00±0.04( P <0.05);与2组比较,培养48、72 h时1组细胞OD值升高( P 均< 0.05 ),与4组比较,培养48、72 h时3组细胞OD值降低( P 均<0.05);培养48 h时1、2组细胞穿膜数分别为73.98± 10.32 、50.67±8.94( P <0.05),3、4组细胞穿膜数分别为28.04±7.99、53.17±10.52( P <0.05);培养48 h时,与2组比较,1组细胞β-catenin、cMYC相对表达量增加( P <0.05),与4组比较,3组细胞β-catenin、cMYC相对表达量降低( P <0.05)。结论 EMs患者异位子宫内膜组织TGF-β 1 mRNA表达升高。加入TGF-β 1的ESCs细胞增殖、侵袭能力增加,β-catenin、cMYC蛋白表达升高;加Objective To observe the expression of transforming growth factor β 1 (TGF-β 1) in ectopic endometrium of patients with endometriosis (EM), to investigate the proliferation and invasion of endometrial stromal cells (ESCs) cultured with TGF-β 1 and its inhibitors, and to explore the possible mechanism. Methods The qRT-PCR was used to detect the expression of TGF-β 1 mRNA in the ectopic endometrial tissues of 100 patients with EM and the normal endometrial tissues of 40 patients without EM. The primary ESCs were isolated, cultured, and identified from fresh tissues of ectopic endometrium. ESCs in the logarithmic phase were divided into groups 1, 2, 3, and 4;ESCs in the groups 1 and 2 were treated with 2 ng/mL of TGF-β 1 factor and 4 μL PBS (using TGF-β 1 factor to prepare solvent), groups 3 and 4 with 1 μmol/L of TGF-β 1 inhibitor Ly364947 and 2 μL DMSO (using Ly364947 to prepare solvent). The expression of TGF-β 1 mRNA was detected by qRT-PCR at 48 h. The proliferation ability (OD) was measured by MTS at 0, 24, 48 and 72 h. The invasive ability was observed by Boyden experiment at 48 h. The expression of β-catenin and cMYC protein was detected by Western blotting at 48 h. Results The relative expression of TGF-β 1 mRNA in the ectopic endometrium and normal endometrium were 2.26±0.96 and 1.02±0.38, respectively ( t =7.908, P <0.05). The relative expression of TGF-β 1 mRNA in the group 1 and group 2 at 48 h was 8.88±1.03 and 1.00±0.02, respectively ( P <0.05), and the relative expression of TGF-β 1 mRNA in the group 3 and group 4 was 0.57±0.07 and 1.00±0.04, respectively ( P <0.05). Compared with the group 2, OD in the group 1 increased at 48 and 72 h ( P <0.05), and OD decreased at 48 and 72 h as compared with that in the group 4 ( P <0.05). At 48 h, the number of transmembrane cells was 73.98±10.32 and 50.67±8.94, respectively, in the group 1 and group 2, versus 28.04±7.99 and 53.17±10.52, respectively, in the group 3 and group 4 (all P <0.05). At 48 h, the relative expression of β-cateni

关 键 词：转化生长因子β1 子宫内膜异位症 子宫内膜间质细胞 细胞增殖 细胞侵袭 Β-连环蛋白 cMYC蛋白 

分 类 号：R711.71[医药卫生—妇产科学]