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. 2017 Jul;19(7):820-825.
doi: 10.7499/j.issn.1008-8830.2017.07.018.

[Effect of annexin A2 on EGFR/NF-κB signal transduction and mucin expression in human airway epithelial cells treated with Mycoplasma pneumoniae]

[Article in Chinese]
Dong-Dong Shen  1 , Fei Yuan, Jiang-Hong Hou
Affiliations

[Effect of annexin A2 on EGFR/NF-κB signal transduction and mucin expression in human airway epithelial cells treated with Mycoplasma pneumoniae]

[Article in Chinese]
Dong-Dong Shen et al. Zhongguo Dang Dai Er Ke Za Zhi. 2017 Jul.

Abstract
in English, Chinese

Objective: To investigate the effect of annexin A2 (AnxA2) on epithelial growth factor receptor (EGFR)/nuclear factor-κB (NF-κB) signal transduction and mucin expression in human airway epithelial H292 cells treated with Mycoplasma pneumoniae (MP).

Methods: H292 cells were divided into control group, MP group, NC-siRNA+MP group, and AnxA2 siRNA+MP group. The cells in the MP group were incubated with 5 μg/mL MP antigen for 2 hours. The cells in the NC-siRNA+MP and AnxA2 siRNA+MP groups were transfected with NC-siRNA and AnxA2 siRNA for 24 hours, followed by MP antigen stimulation for 2 hours. The MTT method was used to measure cell viability; quantitative real-time PCR was used to measure the mRNA expression of AnxA2; Western blot was used to measure the protein expression of AnxA2, phosphorylated EGFR (p-EGFR), and phosphorylated p65 NF-κB (p-p65 NF-κB); ELISA was used to measure the secretion of mucin 5AC (MUC5AC) and mucin 5B (MUC5B).

Results: The MP and NC-siRNA+MP groups had lower cell viability than the control group (P<0.05). The AnxA2 siRNA+MP group had higher cell viability than the MP and NC-siRNA+MP groups and lower cell viability than the control group (P<0.05). The MP and NC-siRNA+MP groups had significantly higher mRNA and protein expression of AnxA2 than the AnxA2 siRNA+MP group (P<0.05). Compared with the control group, the MP and NC-siRNA+MP groups had significant increases in the protein expression of p-EGFR, p-p65 NF-κB, MUC5AC, and MUC5B (P<0.05); the AnxA2 siRNA+MP group had lower protein expression than the MP and NC-siRNA+MP groups, but higher protein expression than the control group (P<0.05).

Conclusions: AnxA2 is involved in the airway lesion induced by MP antigen via mediating EGFR/NF-κB signaling activation and mucin expression in human airway epithelial cells.

目的: 探讨沉默膜联蛋白A2（AnxA2）对肺炎支原体（MP）处理后人气道上皮细胞H292表皮生长因子受体（EGFR）/核因子κB（NF-κB）信号转导及黏蛋白表达的影响。

方法: 将H292细胞分为对照组、MP组、NC-siRNA+MP组和AnxA2 siRNA+MP组。MP组上皮细胞采用5 μg/mL MP抗原孵育2 h。NC-siRNA+MP组和AnxA2 siRNA+MP组细胞分别转染NC-siRNA和AnxA2 siRNA 24 h后用MP抗原刺激2 h。MTT法检测各组细胞活性；实时荧光定量PCR（qRT-PCR）检测各组细胞中AnxA2 mRNA的表达水平；Western blot法检测各组细胞AnxA2、磷酸化EGFR（p-EGFR）、磷酸化p65 NF-κB（p-p65 NF-κB）的表达水平；酶联免疫吸附试验检测黏蛋白5AC（MUC5AC）和黏蛋白5B（MUC5B）的分泌。

结果: MP组和NC-siRNA+MP组细胞活性低于对照组（P < 0.05），AnxA2 siRNA+MP组细胞活性高于MP组和NC-siRNA+MP组，但仍低于对照组（P < 0.05）。MP组和NC-siRNA+MP组AnxA2 mRNA和蛋白表达水平均明显高于AnxA2 siRNA+MP组（P < 0.05）。相比于对照组，MP组和NC-siRNA+MP组中p-EGFR、p-p65 NF-κB、MUC5AC和MUC5B的蛋白表达水平均明显升高（P < 0.05），AnxA2 siRNA+MP组上述蛋白表达水平则低于MP组和NC-siRNA+MP组，但仍高于对照组（P < 0.05）。

结论: AnxA2通过介导人气道上皮细胞EGFR/NF-κB信号活化和黏蛋白表达参与MP抗原诱导的气道病变。

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Figures

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1
MTT法检测各组H292细胞活性（n=3）
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实时荧光定量PCR检测各组H292细胞AnxA2 mRNA表达（n=3）
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Western blot检测各组H292细胞AnxA2蛋白表达
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Western blot检测各组H292细胞p-EGFR和p-p65 NF-κB蛋白表达电泳图
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Western blot检测各组H292细胞p-EGFR和p-p65 NF-κB蛋白表达统计图（n=3）
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