中国生物工程杂志

Objective: To investigate the establishment of the experimental cell model of MUC5AC and MUC5B high expression induced by human neutrophil elastase (HNE) and the relative mechanism. Methods: Human lung adenocarcinoma A549 cells were cultured and divided into six groups：control group, HNE25nM group, HNE50nM group, HNE100nM group, anti-EGFR antibody+HNE50nM group and AG1478+HNE50nM group. The cell activity was assessed by MTT method. The changes of MUC5AC mRNA and MUC5B mRNA were analyzed by RT-PCR. The protein expression changes of MUC5AC and MUC5B were detected by ELISA, while the protein expression changes of MUC5AC, MUC5B and p-EGFR were observed by Immunofluorescence and confocal laser technology. Results: HNE depressed the cell activity in a dose-dependent manner. The gene transcription and the protein expression of MUC5AC and MUC5B in the HNE-induced groups were significantly higher than those in the control group and the differences were statistically significant (P <0.01). p-EGFR protein expression in the HNE50nM group was much more than that in the control group. MUC5AC mRNA and protein in the anti-EGFR antibody+HNE50nM group and the AG1478+HNE50nM group were lower than those in the HNE50nM group, but MUC5B mRNA and protein had no differences between these groups. Conclusion: Human lung adenocarcinoma A549 cells express MUC5AC and MUC5B simultaneously, and HNE can effectively stimulate the high expression of MUC5AC and MUC5B. The A549 cell with mucin high expression induced by HNE can provide the cell model, which is similar with the environment in vivo and helps the research of airway mucus hypersecretion disease. HNE induces the high expression of MUC5AC via EGFR signal transduction pathway, but MUC5B high expression mechanism is different, which need further study.