转化生长因子β1诱导A549细胞上皮-间质转化及其与钙蛋白酶的关系  

Transforming growth factor-β1 induces epithelial-mesenchymal transition in A549 cells by up-regulating expression of calpain

作　　者：谭维军[1] 程真顺[1] 谭秋月[1] 汪婷[1] 连敏[1] 张莉[1] 

机构地区：[1]武汉大学中南医院呼吸内科,430071

出　　处：《中华结核和呼吸杂志》2015年第3期202-205,共4页Chinese Journal of Tuberculosis and Respiratory Diseases

摘　　要：目的 探讨转化生长因子β1（TGF-β1）对肺泡上皮细胞上皮-间质转化的影响及与钙蛋白酶的关系.方法 体外培养人Ⅱ型肺泡上皮细胞来源的A549细胞株,将传代培养的细胞分为空白对照组、TGF-β1处理组、TGF-β1＋钙蛋白酶抑制剂处理组、钙蛋白酶抑制剂组,给予相应处理后Western-blot和RT-PCR分别检测上皮细胞标志物E钙黏素和间质细胞标志物α-平滑肌肌动蛋白（α-SMA）及钙蛋白酶-1（Calpain-1）的表达.结果 与空白对照组对比,TGF-β1组钙蛋白酶-1的mRNA表达及蛋白表达明显增高（P＜0.01）,E钙黏素的mRNA表达及蛋白表达均明显降低（P＜0.01）,α-SMA的mRNA表达及蛋白表达明显增高（P＜0.01）;加入钙蛋白酶抑制剂后再加入TGF-β1刺激,相对于TGF-β1处理组,钙蛋白酶-1的mRNA表达及蛋白表达明显降低（P＜0.01）,E钙黏素的mRNA表达及蛋白表达明显升高（P＜0.05）,α-SMA的mRNA表达水平及蛋白表达水平明显降低（P＜0.01）,而与对照组比较,钙蛋白酶-1、E钙黏素、α-SMA表达无明显差异（P＞0.05）;单加入钙蛋白酶抑制剂后,与空白对照组比较,钙蛋白酶-1、E钙黏素、α-SMA的mRNA表达及蛋白表达无明显差异（P＞0.05）.结论 TGF-β1可以通上调钙蛋白酶的表达诱导A549细胞发生上皮-间质转化.Objective To investigate the effect of transforming growth factor-β1 （TGF-β1） on epithelial-mesenchymal transition in type Ⅱ alveolar epithelial cells and the relationship with Calpain.Methods The type Ⅱ alveolar epithelial cell line-A549 cells cultured in vitro were divided into 4 groups：the control group,the TGF-β1 group,the TGF-β1 ＋ PD150606 group,the PD150606 group.Given the corresponding processing,Western-blot and RT-PCR were utilized to evaluate the expression of Calpain-1,E-cad and α-SMA.Results Compared with the control group,the protein and mRNA expression of Calpain-1 and α-SMA in the TGF-β1 group was markedly increased,but the protein and mRNA expression of E-cad was significantly decreased.While,after calpain inhibitor PD150606 was added,the protein and mRNA expression of Calpain-1 and α-SMA was markedly decreased and the protein and mRNA expression of E-cad was significantly increased in the TGF-β1 ＋ PD150606 group,but there was no difference between TGF-β1 ＋ PD150606 group and control group.But with Calpain inhibitor PD150606 alone,the expression of Calpain-1,E-cad and α-SMA was no difference with the control group.Conclusions TGF-β1 could induce epithelial-mesenchymal transition in A549 cells by up-regulating expression of calpain which could be inhibited by calpain inhibitor PD150606.

关 键 词：肺疾病 间质性 上皮-间质转化 转化生长因子Β1 钙蛋白酶 

分 类 号：R563.9[医药卫生—呼吸系统]