| 摘 要: 目的 观察表皮生长因子受体酪氨酸激酶抑制剂（EGFR-TKI）对二氧化硅（SiO2）所致肺上皮细胞间质转化的影响，并探讨表皮生长因子受体（EGFR）信号通路在肺纤维化上皮间质转化中的作用。方法 体外制备矽肺细胞模型，用EGFR-TKI干预染尘肺上皮细胞，实验分组：正常对照组（N组，SiO2粉尘浓度为0 µg/ml）、模型组（F组，SiO2粉尘浓度为200 µg/ml）、低剂量干预组（A1组，EGFR-TKI浓度20 µmol/L）、中剂量干预组（A2组，EGFR-TKI浓度40 µmol/L）、高剂量干预组（A3组，EGFR-TKI浓度80 µmol/L），干预48 h后，于倒置光学显微镜观察细胞形态学改变，并收集不同时段细胞，用RT-PCR检测 E-cadherin、α-SMA、EGFR mRNA表达水平，细胞免疫荧光方法检测蛋白表达的变化。结果 EGFR-TKI干预，上皮细胞转化停滞，表现为纤维状伪足变短，呈类圆形；E-cadherin mRNA和蛋白表达无明显变化，F组、A1组、A2组、A3组之间差异无统计学意义（P>0.05）；α-SMA mRNA、EGFR mRNA和其蛋白表达逐渐下调，各组之间差异具有统计学意义（P<0.05）。结论 EGFR-TKI能抑制上皮间质转化，其机制可能与下调α-SMA表达和抑制EGFR活性有关。由此推测，抑制EGFR信号通路的活化可能为临床治疗肺纤维化提供新的思路和途径。 |
| 关键词: 表皮生长因子受体酪氨酸激酶抑制剂(EGFR-TKI) 二氧化硅 表皮生长因子受体(EGFR) 上皮间质转化 |
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中图分类号: R994
文献标识码:
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| 基金项目: 2013年青岛市民生计划项目(13-1-3-47-nsh) |
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| Effect of EGFR-TKI on silica-induced epithelial-mesenchymal transition in human A549 cells |
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Zhang Hua, LI Li, Xiao Hua, Zhang Zhunling
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Department of Occupational disease, Qingdao Municipal Central Hospital, Qingdao 266042, China
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| Abstract: Objective To investigate the effect of epidermal growth factor receptor tyrosine kinase inhibitor（EGFR-TKI）on the epithelial-mesenchymal transition (EMT) induced by silica, and explore the role of EGFR signaling pathway in silica-induced EMT in Human A549 cells in vitro. Methods SiO2 exposed lung epithelial cells culture medium added EGFR-TKI, the cells were devided into three groups: control group ( N, SiO2 0μg/ml), silica group (F, SiO2 200 μg/ml), and silica plus EGFR-TKI group (A1, SiO2 200 μg/ml + EGFR-TKI 20μmol/L—low dose subgroup; A2, SiO2 200μg/ml + EGFR-TKI 40μmol/L—middle dose subgroup; A3, SiO2 200μg /ml + EGFR-TKI 80μmol/L—high dose subgroup). After 48h, the cells were observed under phase-constrast microscope, and RT-PCR and immunnoflurescence were used to detect the expression levels of mRNA and protein of E-cadherin,α-SMA, and EGFR. Results , All cells from EGFR-TKI administrated groups showed a stagnation of transforming to mesenchymal phenotype, evidenced by significantly shorter pseudopodia and presented a round shape; the mRNA and protein expression of E-cadherin had no significant change among F group and intervention groups (A1, A2, A3) (P>0.05); while there were some significant differences (P<0.05) among N group and intervention groups (A1, A2, A3) in mRNA and protein expressions ofα-SMA and EGFR . Conclusions The results suggested that EGFR-TKI may inhibit the epithelial-mesenchymal transition by silica dusts, which may be associated with reducedα-SMA expression and inhibition of EGFR activity. Therefore, it is speculated that inhibition of EGFR activation may be a new clue in clinical treatment of pulmonary fibrosis. |
| Keywords: epidermal growth factor receptor tyrosine kinase inhibitor (EGFR-TKI) silica epidermal growth factor receptor (EGFR) epithelial-mesenchymal transition (EMT) |
