Wang Ying-Na, Qin Li, Li Jing-Ming, Chen Li, Pei Cheng
Medical School of Xi'an Jiaotong University, Xi'an 710061, Shaanxi Province, China.
Department of Ophthalmology, the First Affiliated Hospital, Medical School of Xi'an Jiaotong University, Xi'an 710061, Shaanxi Province, China.
Int J Ophthalmol. 2015 Oct 18;8(5):872-6. doi: 10.3980/j.issn.2222-3959.2015.05.04. eCollection 2015.
To investigate the expression of transcription factors Slug in human lens epithelial cells (HLECs) undergoing epithelial-mesenchymal transition (EMT) induced by connective tissue growth factor (CTGF).
HLECs were treated with CTGF of different concentrations (20, 50 and 100 ng/mL) or without CTGF (control) for 24h. The morphological changes of HLECs were analysed by microscopy. The expression and cellular localization of Slug was evaluated by immumo-fluorescence. Expressions of Slug, E-cadherin and alpha smooth muscle actin (α-SMA) were further determined by Western blot analysis.
HLECs showed spidle fibrolasts-like characteristics and loosely connected each other after CTGF treatment. The immuno-fluorescence staining indicated that Slug was localized in the nuclei and its expression was induced by CTGF. The relative expressions of Slug protein were 1.64±0.11, 1.96 ±0.03, 3.12 ±0.10, and 4.08±0.14, respectively, in response to control group and treatment with CTGF of 20, 50 and 100 ng/mL (F=443.86, P<0.01). The increased Slug protein levels were correlated well with up-expression of α-SMA (0.78±0.05, 0.85±0.06, 2.17±0.15, 2.86±0.10; F=449.85, P<0.01) and down-expression of E-cadherin (2.50±0.11, 1.79±0.26, 1.05±0.14, 0.63±0.08; F=101.55, P<0.01).
Transcription factor Slug may be involved in EMT of HLECs induced by CTGF in vitro.
研究转录因子Slug在结缔组织生长因子(CTGF)诱导的人晶状体上皮细胞(HLECs)上皮-间质转化(EMT)过程中的表达。
将HLECs分别用不同浓度(20、50和100 ng/mL)的CTGF处理或不做处理(对照组)24小时。通过显微镜分析HLECs的形态变化。通过免疫荧光评估Slug的表达和细胞定位。通过蛋白质印迹分析进一步测定Slug、E-钙黏蛋白和α平滑肌肌动蛋白(α-SMA)的表达。
CTGF处理后,HLECs呈现出纺锤形成纤维细胞样特征,且细胞间连接松散。免疫荧光染色表明Slug定位于细胞核,其表达由CTGF诱导。对照组以及分别用20、50和100 ng/mL CTGF处理组中,Slug蛋白的相对表达量分别为4.08±0.14、1.64±0.11、1.96±0.03和3.12±0.10(F=443.86,P<0.01)。Slug蛋白水平的升高与α-SMA的上调(0.78±0.05、0.85±0.06、2.17±0.15、2.86±0.10;F=449.85,P<0.01)和E-钙黏蛋白的下调(2.50±0.11、1.79±0.26、1.05±0.14、C0.63±0.08;F=101.55,P<0.01)密切相关。
转录因子Slug可能参与体外CTGF诱导的HLECs的EMT过程。