Integrated analysis of DNA methylation and mRNA expression profiling reveals candidate genes associated with cisplatin resistance in non-small cell lung cancer.
DNA methylation plays a critical role during the development of acquired chemoresistance. The aim of this study was to identify candidate DNA methylation drivers of cisplatin (DDP) resistance in non-small cell lung cancer (NSCLC). The A549/DDP cell line was established by continuous exposure of A549 cells to increasing concentrations of DDP. Gene expression and methylation profiling were determined by high-throughput microarrays. Relationship of methylation status and DDP response was validated in primary tumor cell culture and the Cancer Genome Atlas (TCGA) samples. Cell proliferation, apoptosis, cell cycle, and response to DDP were determined in vitro and in vivo. A total of 372 genes showed hypermethylation and downregulation in A549/DDP cells, and these genes were involved in most fundamental biological processes. Ten candidate genes (S100P, GDA, WISP2, LOXL1, TIMP4, ICAM1, CLMP, HSP8, GAS1, BMP2) were selected, and exhibited varying degrees of association with DDP resistance. Low dose combination of 5-aza-2'-deoxycytidine (5-Aza-dC) and trichostatin A (TSA) reversed drug resistance of A549/DDP cells in vitro and in vivo, along with demethylation and restoration of expression of candidate genes (GAS1, TIMP4, ICAM1 and WISP2). Forced expression of GAS1 in A549/DDP cells by gene transfection contributed to increased sensitivity to DDP, proliferation inhibition, cell cycle arrest, apoptosis enhancement, and in vivo growth retardation. Together, our study demonstrated that a panel of candidate genes downregulated by DNA methylation induced DDP resistance in NSCLC, and showed that epigenetic therapy resensitized cells to DDP.
Department of Oncology; Jinling Hospital; Medical School of Nanjing University; Nanjing, PR China; Department of Oncology; Yangzhou No. 1 Hospital; The Second Clinical School of Yangzhou University; Yangzhou, PR China; Department of Oncology; Affiliated Xuzhou Central Hospital; Xuzhou Medical College; Xuzhou, PR China.
You-Wei
YW
Zhang
Department of Oncology; Jinling Hospital; Medical School of Nanjing University; Nanjing, PR China.
Yun
Y
Zheng
Department of Oncology; Jinling Hospital; Medical School of Nanjing University; Nanjing, PR China.
Jing-Zi
JZ
Wang
Department of Oncology; Yangzhou No. 1 Hospital; The Second Clinical School of Yangzhou University; Yangzhou, PR China.
Xiao-Xia
XX
Lu
Department of Oncology; Yangzhou No. 1 Hospital; The Second Clinical School of Yangzhou University; Yangzhou, PR China.
Zhu
Z
Wang
Department of Oncology; Jinling Hospital; Medical School of Nanjing University; Nanjing, PR China.
Long-Bang
LB
Chen
Department of Oncology; Jinling Hospital; Medical School of Nanjing University; Nanjing, PR China.
Xiao-Xiang
XX
Guan
Department of Oncology; Yangzhou No. 1 Hospital; The Second Clinical School of Yangzhou University; Yangzhou, PR China.
Jian-Dong
JD
Tong
D000964:Antimetabolites, Antineoplastic / Q000494:pharmacology; D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis; D001374:Azacitidine / Q000031:analogs & derivatives / Q000494:pharmacology; D002289:Carcinoma, Non-Small-Cell Lung / Q000235:genetics* / Q000473:pathology; D059447:Cell Cycle Checkpoints; D018797:Cell Cycle Proteins / Q000378:metabolism; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D002945:Cisplatin / Q000494:pharmacology*; D019175:DNA Methylation; D000077209:Decitabine; D019008:Drug Resistance, Neoplasm / Q000187:drug effects*; D044127:Epigenesis, Genetic; D058851:GPI-Linked Proteins / Q000378:metabolism; D020869:Gene Expression Profiling / Q000379:methods; D056572:Histone Deacetylase Inhibitors / Q000494:pharmacology; D006801:Humans; D006877:Hydroxamic Acids / Q000494:pharmacology; D008175:Lung Neoplasms / Q000235:genetics* / Q000473:pathology; D012333:RNA, Messenger / Q000378:metabolism*
D000964:Antimetabolites, Antineoplastic; D000970:Antineoplastic Agents; D018797:Cell Cycle Proteins; C096192:GAS1 protein, human; D058851:GPI-Linked Proteins; D056572:Histone Deacetylase Inhibitors; D006877:Hydroxamic Acids; D012333:RNA, Messenger; C012589:trichostatin A; D000077209:Decitabine; D001374:Azacitidine; D002945:Cisplatin
5-aza-2′-deoxycytidine; GAS1; cisplatin; lung cancer; methylation; trichostatin A