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Effect of histone acetylation on asthma susceptibility gene ADAM33 in A549 and BEAS-2B cells |
HAO Zhongfen 1, DU Juan 1, RUI Feifei 2, HANG Yun 1, TANG Chenlu 1, LI Zhang 1, SHU Jin 1 |
(1. Department of Pediatrics, the Fourth Affiliated Hospital of Jiangsu University, Zhenjiang Jiangsu 212001; 2. Department of Neonatal, Changzhou Maternal and Child Health Hospital, Changzhou Jiangsu 213000, China) |
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Abstract Objective: To evaluate the effects of histone acetylation mediated by sodium butyrate, trichostatin A (TSA) and adenoviral E1A binding protein of 300 kD (P300) on the expression of a disintegrin and metalloproteinase 33 (ADAM33), one of asthma susceptible genes, in human non-small cell lung cancer A549 cells and human bronchial epithelial BEAS-2B cells. Methods: Human non-small cell lung cancer A549 cells and human bronchial epithelial BEAS-2B cells were divided into sodium butyrate control group (double distilled water treatment) and 1, 2.5, 5 mmol/L sodium butyrate groups, TSA control group (0.1% dimethyl sulfoxide treatment) and 0.2, 0.4, 0.8 μmol/L TSA groups, respectively. BEAS-2B cells were divided into P300 control group (transfected with P300 mutant plasmid) and P300 group (transfected with P300 expressing plasmid). These cells were treated respectively according to the groups. The ADAM33 promoter activity was analyzed by dual luciferase reporter method, the expression of ADAM33 mRNA was detected by quantitative real time-PCR (qRT-PCR), and the expression of ADAM33 protein was detected by Western blotting. Results: In human non-small cell lung cancer A549 cells, compared with the control group, ADAM33 promoter activity was significantly decreased in 1 mmol/L sodium butyrate group and 0.2 μmol/L TSA group (P<0.01). In human bronchial epithelial BEAS-2B cells, compared with the control group, the activity of ADAM33 gene promoter, mRNA and protein expression levels was greatly decreased in 1 mmol/L sodium butyrate group and 0.2 μmol/L TSA group (P<0.05 or P<0.01). There was no significant difference in ADAM33 expression when increasing the concentration of sodium butyrate and TSA. In BEAS-2B cells, compared with the control group, the promoter activity, mRNA and protein expression levels of ADAM33 in P300 group were markedly decreased (P<0.05). Conclusion: Sodium butyrate and TSA decreased ADAM33 expression in human non-small cell lung cancer A549 cells and human bronchial epithelial BEAS-2B cells through histone acetylation, while P300 decreased ADAM33 expression in human bronchial epithelial BEAS-2B cells through histone acetylation.
[Key words]asthma; a disintegrin and metalloproteinase 33 (ADAM33); sodium butyrate; trichostatin A; human bronchial epithelial BEAS-2B cells
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Received: 10 January 2023
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