Overexpression of DNA damage-induced 45 α gene contributes to esophageal squamous cell cancer by promoter hypomethylation

Bao Xiang Wang,Xin Min Zhou,Chen Chen,Jing Qun Tang,Bang Liang Yin,Ni Yin,Yi Zhi Pan,Zhen Kun Xia,Bin He,Ming Zhao,Wei Xing Zhang

doi:10.1186/1756-9966-31-11

Abstract

BackgroundEnvironmental factors-induced dysfunction of esophageal squamous epithelium, including genomic DNA impairment and apoptosis, play an important role in the pathogenesis of esophageal squamous cell cancer. DNA damage-induced 45α (GADD45α) has been found promoting DNA repair and removing methylation marker, Therefore, in this study we will investigate whether GADD45α expression is induced and its mechanism in esophageal squamous cell cancer.MethodsTwo human esophageal squamous cell lines (ESCC), ECA109 and KYSE510 were cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum (FBS). Lipofectamine 2000 was used to transfect cells. mRNA level of GADD45α was measured by reverse transcription-quantitive PCR (RT-qPCR), protein level of GADD45α was detected by western blot and Immunohistochemistry. Global DNA methylation of tissue sample was measured using the Methylamp Global DNA Methylation Quantification Ultra kit (Epigentek Group) and promoter methylation was measured by bisulfite sequencing.ResultsGADD45a mRNA and protein levels were increased significantly in tumor tissue than that in adjacent normal tissue. Hypomethylation of global genomic DNA and GADD45α promoter were found in ESCC. The cell sensitivity to Cisplatin DDP was decreased significantly in Eca109 and Kyse510 cells, in which GADD45α expression was down-regulated by RNA interference (RNAi). In addition, silence of GADD45a expression in ESCC cells inhibited proliferation and promoted apoptosis.ConclusionOverexpression of GADD45α gene is due to DNA hypomethylation in ESCC. GADD45α may be a protective factor in DDP chemotherapy for esophageal squamous cell carcinoma.

Highlights

Environmental factors-induced dysfunction of esophageal squamous epithelium, including genomic DNA impairment and apoptosis, play an important role in the pathogenesis of esophageal squamous cell cancer
Mice lacking the Growth arrest and DNA damage-induced 45a (GADD45a) gene are susceptible to DNA damage-induced tumors, including carcinogenesis induced by ionizing radiation, UV radiation and dimethylbenzanthracene (DMBA) [17,18]
GADD45a mRNA level was higher in tumor tissue than in adjacent normal tissue (P = 0.001) (Figure 1A and Table 3)

Summary

Introduction

Environmental factors-induced dysfunction of esophageal squamous epithelium, including genomic DNA impairment and apoptosis, play an important role in the pathogenesis of esophageal squamous cell cancer. DNA damage-induced 45a (GADD45a) has been found promoting DNA repair and removing methylation marker, in this study we will investigate whether GADD45a expression is induced and its mechanism in esophageal squamous cell cancer. Esophageal squamous cell carcinoma (ESCC) is the predominant type of esophageal cancer, comprising almost 95% of cases. Mice lacking the GADD45a gene are susceptible to DNA damage-induced tumors, including carcinogenesis induced by ionizing radiation, UV radiation and dimethylbenzanthracene (DMBA) [17,18]. A recent study showed that GADD45a has a key role in active DNA demethylation and its overexpression activates methylation-silenced reporter plasmids and promotes global DNA demethylation. A recent study showed that GADD45a has a key role in active DNA demethylation and its overexpression activates methylation-silenced reporter plasmids and promotes global DNA demethylation. [19]

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Conclusion