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Hypermethylation of ATP ‐binding cassette B1 ( ABCB 1 ) multidrug resistance 1 ( MDR 1 ) is associated with cisplatin resistance in the A549 lung adenocarcinoma cell line
Author(s) -
Li Angui,
Song Jianfei,
Lai Qi,
Liu Bangqing,
Wang Haiyong,
Xu Yinhui,
Feng Xiaoyan,
Sun Xiaolin,
Du Zhenzong
Publication year - 2016
Publication title -
international journal of experimental pathology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.671
H-Index - 72
eISSN - 1365-2613
pISSN - 0959-9673
DOI - 10.1111/iep.12212
Subject(s) - cisplatin , a549 cell , microbiology and biotechnology , methylation , cancer research , lung cancer , gene expression , biology , dna methylation , adenocarcinoma , multiple drug resistance , apoptosis , gene , cancer , drug resistance , pathology , medicine , chemotherapy , genetics
Summary Development of multiple drug resistance has been attributed to the overexpression of the ATP ‐binding cassette B1 ( ABCB 1 ) gene. In this study, the major purpose was to assess the expression and methylation levels of ABCB 1 in human lung adenocarcinoma and to reveal the relationship between these processes and acquisition of cisplatin ( DDP ) resistance in the human cancer cell line A549. Methylation and expression levels of the ABCB 1 gene ABCB 1 in clinical human lung tissue were assessed using bisulphite sequencing, reverse transcription real‐time PCR ( RT 2 ‐ PCR ) and Western blot methods. Cell viability, DDP resistance and apoptosis of A549 cells were evaluated using the Cell Counting Kit‐8 and fluorescence‐activated cell sorter analysis. Our results showed that the onset of resistance to the cisplatin analogue, DDP , was associated with hypermethylation of the ABCB 1 gene. Expression of the ABCB 1 gene was enhanced at both mRNA and protein levels. Treatment with 5‐Aza‐C contributed to the hypomethylation of the ABCB 1 gene and decreased ABCB 1 protein expression in A549 cells. In conclusion, this in vitro and human tissue study of lung adenocarcinoma cells demonstrated that hypermethylation of the ABCB 1 gene correlated with increased gene expression and was associated with the acquisition of resistance to the cisplatin analogue, DDP in human lung adenocarcinoma cells. Taken together, our study highlighted the connection between increased ABCB 1 methylation level and upregulated expression of the gene in lung cancer. Moreover, the abnormally high expression of ABCB 1 in A549 cells contributed to the development of the DDP resistance.