HMGB1 RNAi抑制TGF-β1诱导的A549细胞株EMT

基础医学与临床 ›› 2015, Vol. 35 ›› Issue (2): 183-186.

HMGB1 RNAi抑制TGF-β1诱导的A549细胞株EMT

蔡琳¹,阮志燕¹,徐军²

1. 广东食品药品职业学院
2. 广州医学院第一附属医院呼吸内科

收稿日期:2014-06-16 修回日期:2014-10-06 出版日期:2015-02-05 发布日期:2015-01-23
通讯作者: 蔡琳 E-mail:lin_cai@126.com
基金资助:
广东省医学科研基金

HMGB1 RNAi inhibits TGF-β1 induced epithelial-mesenchymal transition in A549 cells

Received:2014-06-16 Revised:2014-10-06 Online:2015-02-05 Published:2015-01-23

摘要/Abstract

摘要： 目的探讨HMGB1在肺泡上皮细胞间质转分化中的作用。方法设计并合成针对HMGB1的小干扰RNA(siRNA)，体外培养Ⅱ型肺泡上皮细胞系细胞-A549细胞，将细胞分为4组：1）对照组，2）TGF-β1刺激组，3）RNAi组（TGF-β1+ HMGB1 siRNA），4）RNAi阴性对照组（TGF-β1+ siRNA阴性对照）；倒置相差显微镜观察细胞形态学；RT-PCR法检测HMGB1和α平滑肌肌动蛋白（α-SMA）的mRNA表达；Western blot法检测α-SMA和HMGB1蛋白表达。结果 TGF-β1刺激组细胞HMGB1和α-SMA 的mRNA和蛋白表达均较对照组显著增加(P＜0.01)；RNAi组HMGB1和α-SMA mRNA和蛋白表达均较正常对照组组显著下降(P＜0.01)。结论 HMGB1可能参与了TGFβ1 诱导的肺泡上皮细胞转分化。

关键词: 关键词：RNA干扰, 高迁移率族蛋白B1, α平滑肌肌动蛋白, 上皮间质转分化

Abstract: Objective To investigate the role of HMGB1 in epithelial-mesenchymal transition. Methods Specific siRNA of HMGB1 were designed and synthesized. Cultured typeⅡ alveolar epithelial cell line-A549 cells were divided into 4 groups: (1) control group, (2) model group induced by TGF-β1, (3) HMGB1 RNAi group, (4) RNAi negative control group. Cellular morphology changes were observed by phase-contrast microscope. HMGB1 and α-SMA expression in A549 cells was detected by RT-PCR and Western blotting respectively. Results mRNA and protein expression of HMGB1 and α-SMA in TGF-β1 group increased significantly than that in normal control group(P＜0.01). Both of HMGB1 and α-SMA mRNA and protein expression in siRNA-treated cells were decreased significantly compared with that in TGF-β1 group (P<0.01). Conclusions HMGB1 may participate in the TGF- β 1 induced EMT．

Key words: Key words: RNAi, High mobility group B1, α-smooth muscle actin, epithelial-mesenchymal transition

中图分类号:

R563

蔡琳阮志燕徐军. HMGB1 RNAi抑制TGF-β1诱导的A549细胞株EMT[J]. 基础医学与临床, 2015, 35(2): 183-186.