转化生长因子-β2诱导的上皮间质转分化对人晶状体上皮细胞基因表型的影响

Effects of transforming growth factor-β2 induced epithelial-mesenchcymal transition in human lens epithelial cells on expression of its gene phenotype

目的观察转化生长因子-β2(TGF-β2)诱导人晶状体上皮细胞(LECs)发生上皮间质转分化(EMT)时,其形态和基因表型所发生的变化。方法在体外培养的人LECs中,加入100 pg/mL浓度的TGF-β2,诱导其发生EMT,处理0、6、24、48 h后,观察其在形态上的变化,利用实时定量反转录聚合酶链反应(qRTPCR)于各时间点检测其标志性基因E-钙粘蛋白(CDH1)、α-平滑肌肌动蛋白(α-SMA)、波形蛋白(VIM)等在表达水平上所发生的变化,并与未处理的空白对照组进行比较。结果加入TGF-β2后,随着时间的推移,人LECs在形态上拉长,呈梭形;上皮细胞标志性基因如CDH1的表达水平逐渐下调,48 h时相对表达量为1.10E-06,和对照组2.68E-06相比差异有统计学意义(P=0.002);间质细胞标志性基因如VIM的表达水平逐渐上调,48 h时相对表达量为1.64E-02,和对照组1.37E-02相比差异有统计学意义(P=0.006);而α-SMA的表达水平也逐渐上升,48 h时相对表达量为9.21E-03,虽和对照组8.67E-03相比差异无统计学意义(P=0.551),但仍呈上升趋势。结论在TGF-β2诱导下,人LECs发生EMT,其形态和基因表型均发生与间质细胞相符的变化。

Objective To explore the affects of transforming growth factor-β2 induced epithelial-mesenchyme transition（EMT） in human lens epithelial cells （LECs）Q.n its morphology and gene expression. Methods LECs were cultured in vitro and treated with TGF-β2 at the concentration of 100 pg/mL. The morphology of human LECs were observed and QPCR was used to detect the expression of its gene. Results After 0, 6, 24, 48 h treating with TGF-β2, human LECs were elongated to become a spindle shape with the time. At each time point, the results of QPCR indicated that the epithelial marker genes, such as E-cadherin（ CDH1 ）, was down-regulated, meanwhile, the mesenchymal marker genes, such as vimentin （VIM）, was up-regulated, and or-smooth muscle actin （ot-SMA） expression levels were increased in human LECs. Compared with the control group, the difference of CDH1 and VIM were significantly different （P 〈 0.05 ）, but the difference of expression level of α-SMA was not statistically significant （P 〉 0.05 ）. Conclusions EMT of human LECs could be induced by TGF-β2. The gene expression and morphology changes are inaccordance with mesenchymal ceils.