American Journal of Clinical and Experimental Medicine
Volume 7, Issue 2, March 2019, Pages: 54-60
Received: May 29, 2019;
Published: Jul. 29, 2019
Views 144 Downloads 15
Tingting Chen, Department of Gastroenterology, Binzhou People's Hospital, Binzhou City, P. R. China
Donghui Tian, Department of Neurology, Binzhou People's Hospital, Binzhou City, P. R. China
Yurong Zhang, Department of Hepatobiliary Surgery, Binzhou People's Hospital, Binzhou City, P. R. China
Objective: To identify the methylation silenced tumor suppressor genes in Epstein-Barr virus (EBV)-associated gastric cancer (EBVaGC). Methods: EBV-positive (GT38, PT and SNU719) and negative (SGC7901) gastric cancer cell lines were selected and treated with 5-Aza-CdR. Then real-time fluorescence quantitative PCR was used to validate the results of microarray, and methylation-specific PCR (MSP) and bisulfite genomic sequencing (BGS) were adopted to detect the CpG island methylation levels of gene promoters. Results: The expression levels of 6 differentially expressed genes (H19, LOXL1, ARMCX2, LXN, CDH3 and MMP7) before and after 5-Aza-CdR treatment were confirmed by real-time qPCR, which are consistent with the results of microarray analysis. There were different degrees of methylation in LOXL1 gene promoter in EBVaGC. GT38 and PT were fully methylated, and SGC7901 and HGC-27 was unmethylated, suggesting that this gene is a candidate methylation silenced tumor suppressor gene. The methylation rate of LOXL1 in EBVaGC was significantly higher than that in EBV-negative gastric cancer (EBVnGC). Conclusion: The promoter region of candidate tumor suppressor gene LOXL1 shows high methylation status, indicating that EBV critically accounts for the methylation of LOXL1 gene regulatory region. EBV is involved in the pathogensis of EBVaGC that aberrant methylation occurred in promoter CpG island, which inactivates tumor suppressor genes.
Methylation of Epstein-Barr Virus-Associated Gastric Cancer Suppressor Genes, American Journal of Clinical and Experimental Medicine.
Vol. 7, No. 2,
2019, pp. 54-60.
Young KH, Zhang D, Malik JT, Williams EC. Fulminant EBV-driven CD8 T-cell lymphoproliferative disorder following primary acute EBV infection: a unique spectrum of T-cell malignancy. Int J Clin Exp Pathol 2008, 1: 185-197.
Katsumura KR, Maruo S, Takada K. EBV lytic infection enhances transformation of B-lymphocytes infected with EBV in the presence of T-lymphocytes. J Med Virol 2012, 84: 504-510.
Shamaa AA, Zyada MM, Wagner M, Awad SS, Osman MM, Abdel Azeem AA. The significance of Epstein Barr virus (EBV) & DNA topoisomerase II alpha (DNA-Topo II alpha) immunoreactivity in normal oral mucosa, oral epithelial dysplasia (OED) and oral squamous cell carcinoma (OSCC). Diagn Pathol 2008, 3: 45.
Long HM, Leese AM, Chagoury OL, Connerty SR, Quarcoopome J, Quinn LL, et al. Cytotoxic CD4+ T cell responses to EBV contrast with CD8 responses in breadth of lytic cycle antigen choice and in lytic cycle recognition. J Immunol 2011, 187: 92-101.
Lee SM, Oh YL, Kim KM, Ko YH, Lee J. Coexistence of EBV associated nasopharyngeal undifferentiated carcinoma and gastric lymphoepithelioma-like carcinoma. Pathology 2010, 42: 684-686.
Rusiecki JA, Al-Nabhani M, Tarantini L, Chen L, Baccarelli A, Al-Moundhri MS. Global DNA methylation and tumor suppressor gene promoter methylation and gastric cancer risk in an Omani Arab population. Epigenomics 2011, 3: 417-429.
Li L, Su X, Choi GC, Cao Y, Ambinder RF, Tao Q. Methylation profiling of Epstein-Barr virus immediate-early gene promoters, BZLF1 and BRLF1 in tumors of epithelial, NK- and B-cell origins. BMC Cancer 2012, 12: 125.
Zhao C, Bu X. Promoter methylation of tumor-related genes in gastric carcinogenesis. Histol Histopathol 2012, 27: 1271-1282.
Grafodatskaya D, Choufani S, Ferreira JC, Butcher DT, Lou Y, Zhao C, et al. EBV transformation and cell culturing destabilizes DNA methylation in human lymphoblastoid cell lines. Genomics 2010, 95: 73-83.
Jin Q, Liu C, Yan C, Tao B, Li Z, Cai Z. 5-aza-CdR induces the demethylation of Syk promoter in nasopharyngeal carcinoma cell. Gene 2012, 511: 224-226.
Liu X, Gao Y, Luo B, Zhao Y. Construction and Antiapoptosis Activities of Recombinant Adenoviral Expression Vector Carrying EBV Latent Membrane Protein 2A. Gastroenterol Res Pract 2011, 2011: 182832.
Chen JN, He D, Tang F, Shao CK. Epstein-Barr virus-associated gastric carcinoma: a newly defined entity. J Clin Gastroenterol 2012, 46: 262-271.
Bengtson P, Zetterberg H, Mellberg T, Pahlsson P, Larson G. Characterization of EBV-transformed B-cells established from an individual homozygously mutated (G329A) in the FUT7 alpha1,3-fucosyltransferase gene. Scand J Immunol 2005, 62: 251-258.
Seo JS, Jun SM, Kwon SW, Oh IH, Kim TG, Lee SK. Establishment and characterization of gastric carcinoma cell clones expressing LMP2A of Epstein-Barr virus. Int J Mol Med 2010, 25: 11-16.
Lee JH, Kim SH, Han SH, An JS, Lee ES, Kim YS. Clinicopathological and molecular characteristics of Epstein-Barr virus-associated gastric carcinoma: a meta-analysis. J Gastroenterol Hepatol 2009, 24: 354-365.
Sun Y, Jiang X, Xu Y, Ayrapetov MK, Moreau LA, Whetstine JR, et al. Histone H3 methylation links DNA damage detection to activation of the tumour suppressor Tip60. Nat Cell Biol 2009, 11: 1376-1382.
Ren JZ, Huo JR. [Effect of 5-Aza-CdR on expression and methylation of E-cadherin gene in human colon carcinoma cells]. Chin J Cancer 2010, 29: 38-42.
Painter HJ, Altenhofen LM, Kafsack BF, Llinas M. Whole-genome analysis of Plasmodium spp. Utilizing a new agilent technologies DNA microarray platform. Methods Mol Biol 2013, 923: 213-219.
Naher L, Kiyoshima T, Kobayashi I, Wada H, Nagata K, Fujiwara H, et al. STAT3 signal transduction through interleukin-22 in oral squamous cell carcinoma. Int J Oncol 2012, 41: 1577-1586.
Zhao J, Jin H, Cheung KF, Tong JH, Zhang S, Go MY, et al. Zinc finger E-box binding factor 1 plays a central role in regulating Epstein-Barr virus (EBV) latent-lytic switch and acts as a therapeutic target in EBV-associated gastric cancer. Cancer 2012, 118: 924-936.
Bell A, Bell D, Weber RS, El-Naggar AK. CpG island methylation profiling in human salivary gland adenoid cystic carcinoma. Cancer 2011, 117: 2898-2909.
Liu X, Tang X, Zhang S, Wang Y, Wang X, Zhao C, et al. Methylation and Expression of Retinoblastoma and Transforming Growth Factor-beta1 Genes in Epstein-Barr Virus-Associated and -Negative Gastric Carcinomas. Gastroenterol Res Pract 2012, 2012: 906017.
Sahin M, Sahin E, Gumuslu S, Erdogan A, Gultekin M. DNA methylation or histone modification status in metastasis and angiogenesis-related genes: a new hypothesis on usage of DNMT inhibitors and S-adenosylmethionine for genome stability. Cancer Metastasis Rev 2010, 29: 655-676.
Gravina GL, Marampon F, Piccolella M, Motta M, Ventura L, Pomante R, et al. Hormonal therapy promotes hormone-resistant phenotype by increasing DNMT activity and expression in prostate cancer models. Endocrinology 2011, 152: 4550-4561.
Park SY, Kook MC, Kim YW, Cho NY, Kim TY, Kang GH. Mixed-type gastric cancer and its association with high-frequency CpG island hypermethylation. Virchows Arch 2010, 456: 625-633.
Perera RA, Samaranayake LP, Tsang CS. Shedding dynamics of Epstein-Barr virus: A type 1 carcinogen. Arch Oral Biol 2010, 55: 639-647.
Engels N, Yigit G, Emmerich CH, Czesnik D, Schild D, Wienands J. Epstein-Barr virus LMP2A signaling in statu nascendi mimics a B cell antigen receptor-like activation signal. Cell Commun Signal 2012, 10: 9.
Ushiku T, Chong JM, Uozaki H, Hino R, Chang MS, Sudo M, et al. p73 gene promoter methylation in Epstein-Barr virus-associated gastric carcinoma. Int J Cancer 2007, 120: 60-66.