摘要
目的:观察TGF-β1作用下,肺泡Ⅱ型上皮细胞RLE-6TN在EMT过程中HDAC1、8的表达情况及TSA对TGF-β1诱导细胞EMT的影响。方法:将TGF-β1加入到体外培养的细胞中,于不同时间收取细胞,采用WB及Real-time RT-PCR检测HDAC1、8及E-cad、α-SMA的表达情况。然后将TGF-β1加入经过TSA预处理的细胞中,于不同时间收取细胞,重新检测上述基因。结果:加入TGF-β1后,E-cad蛋白表达下调;α-SMA蛋白表达上调;HDAC1蛋白表达上调;HDAC8蛋白表达下调。E-cad mRNA表达下调;α-SMA及HDAC8的mRNA均于12 h表达上调,6 h、24 h表达下调;HDAC1 mRNA表达于6 h下调,24 h上调。加入TSA后,E-cad蛋白表达上调;α-SMA、HDAC1及HDAC8蛋白的表达均下调。E-cad mRNA表达上调;α-SMA mRNA表达于6 h、12 h上调,24 h下调;HDAC1 mRNA表达下调;HDAC8 mRNA表达上调。结论:TGF-β1体外诱导的RLE-6TN细胞EMT,可以通过应用TSA抑制其获得α-SMA表型、失去E-cad表型,从而部分逆转TGF-β1诱导的肺泡Ⅱ型上皮EMT。
Objective： To observe the expression of histone deacetylase 1,8 on the process of TGF-β 1 induced type II alveolar epithelial cells to EMT, and investigate the effect of TSA on TGF-β1-induced the cells EMT process. Methods： RLE-6TN cell lines were cultured and treated with TGF- β 1, and collected cells at different time points. The expression of markers and mRNA of E-cad,α-SMA,HDAC1 and HDAC8 were assayed by Western Blot and Real-time RT-PCR, respectively. Then add TGF- β 1 to the cells after TSA pretreatment for 6 hours and repeat the above assay. Results： With the treatment of TGF- β 1, the expression of E-cad protein was down regulated;α -SMA, HDAC1 and HDAC8 protein was unregulated. E-cad mRNA was down regulated; both the mRNA of ct -SMA and HDAC8 were up regulated at 12 h, and were down regulated at 6 h, 24 h; HDAC1 mRNA at 6 h were down regulated, 24 h was up regulated. After pretreated with TSA, the expression of E-cad protein was up regulated; α -SMA, HDAC 1 and HDAC8 protein was down regulated. E-cad mRNA was up regulated; α-SMA mRNA at 6 h, 12 h was up regulated, 24 h was down regulated; HDAC1 mRNA was down regulated; HDAC8 mRNA was up regulated. Conclusion： It is possible to use TSA inhibited RLE-6TN cells to receive α-SMA phenotype, loss of E-cad phenotype, which partially reverse the EMT of RLE-6TN induced by TGF- 13 1 in vitro.
出处
《大理学院学报（综合版）》
CAS
2013年第3期16-20,共5页
Journal of Dali University
基金
国家自然科学基金青年科学基金资助项目(81100045)
