GALC gene is downregulated by promoter hypermethylation in Epstein-Barr virus-associated nasopharyngeal carcinoma

Zhao,Yinghai; Guo,Ying; Wang,Zhijing; Xiao,Zebin; Li,Rong; Luo,Aihua; Wu,Changli; Jing,Zhiliang; Sun,Ning; Chen,Xiaoyi; Du,Haijun; Zeng,Yi

doi:10.3892/or.2015.4134

GALC gene is downregulated by promoter hypermethylation in Epstein-Barr virus-associated nasopharyngeal carcinoma

Authors:
- Yinghai Zhao
- Ying Guo
- Zhijing Wang
- Zebin Xiao
- Rong Li
- Aihua Luo
- Changli Wu
- Zhiliang Jing
- Ning Sun
- Xiaoyi Chen
- Haijun Du
- Yi Zeng
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Affiliations: Department of Pathology, Guangdong Medical University, Zhanjiang, Guangdong 524023, P.R. China, Department of Pathophysiology, Guangdong Medical University, Zhanjiang, Guangdong 524023, P.R. China, Department of Pathology, Gaozhou People's Hospital, Gaozhou, Guangdong 525200, P.R. China, Department of Physiology, Guangdong Medical University, Zhanjiang, Guangdong 524023, P.R. China, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Prevention and Control, State Key Laboratory for Infectious Disease Prevention and Control, Beijing 100052, P.R. China
Published online on: July 14, 2015 https://doi.org/10.3892/or.2015.4134
Pages: 1369-1378

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Abstract

The aim of the study was to investigate the tumor-suppressor effect of GALC in Epstein-Barr virus (EBV)-associated nasopharyngeal carcinoma (NPC) and determine whether GALC was downregulated by promoter hypermethylation in the NPC cell line, CNE-2Z. Forty-one archival NPC biopsy specimens were compared with 15 chronic nasopharyngitis specimens. EBV-encoded RNA (EBER) was verified by in situ hybridization and GALC protein expression was analyzed by immunohistochemistry. Promoter methylation in CNE-2Z cells was analyzed by bisulfite sequencing polymerase chain reaction. The functional role of GALC in NPC was investigated by restoring GALC expression in CNE-2Z cells via treatment with the DNA-demethylating agent 5-Aza-2'-deoxycytidine (5-Aza‑dC). EBER was expressed in 92.68% NPC specimens but no chronic nasopharyngitis specimens (P<0.01). GALC protein was present in 60% of chronic nasopharyngitis specimens and 24.39% NPC specimens (P<0.05). GALC protein expression was present significantly more frequently in tumors without lymph node metastasis than in those with metastasis (P<0.05). Logistic regression showed that GALC protein expression protected against lymph node metastasis (P<0.05). GALC protein expression was not correlated with age, gender and TNM stage (P>0.05). Treatment of GALC-negative CNE-2Z cells with 5-Aza-dC reduced GALC promoter methylation and restored GALC expression in a dose-dependent manner (P<0.05). The re-expression of GALC in CNE-2Z cells reduced cell proliferation and migration compared to the controls (P<0.05). GALC was downregulated by promoter hypermethylation and contributed to the pathogenesis of EBV-associated NPC. The findings showed the putative tumor-suppressor effect of GALC in NPC.

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