Gastric cancer is an aggressive disease which is the fourth prevalent malignancy in the world. Beside the genetic factors, epigenetic alterations such as promoter CpG island hyper methylation are involved in the emergence of gastric cancer. Herein, we investigated the methylation status of CDH11, EphA5, and HS3ST2 genes in patients with and without gastric adenocarcinoma for the first time.
In the study 40 paraffin-embedded tissue sections from gastric adenocarcinoma patients and 40 specimens from patients with functional dyspepsia were taken. DNA extraction was performed using a modified salting out method. Epizen DNA methylation kit was used to the bisulfite DNA conversion. The methylation status of CDH11, EphA5, and HS3ST2 genes were analyzed by methylation-specific PCR (MSP) technique.
Among the 80 specimens, 71 DNA samples were achieved (34 gastric adenocarcinoma patients and 37 control patients). The results showed that CDH11, EphA5, and HS3ST2 genes are methylated in 28 (82.45%), 19 (55.88%), and 26 (76.47%) of 34 DNA samples from gastric adenocarcinoma patients, respectively, whereas, these genes are methylated in 7 (18.91%), 9 (24.32%) and 7 (18.91%) of 37 samples from noncancerous patients, respectively. Statistical analyses using a chi-squared test showed that there is a statistically significant difference in methylation level of CDH11, EphA5, and HS3ST2 genes between gastric cancer and uncancerous patients (p < 0.05).
To the best of our knowledge, this is the first report on methylation of CDH11, EphA5, and HS3ST2 promoters’ in gastric adenocarcinoma patients using MSP. Identification of novel cancer-related molecular mechanisms can be useful in detection of new treatment strategies.
This is a preview of subscription content, access via your institution.
Buy single article
Instant access to the full article PDF.
Tax calculation will be finalised during checkout.
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
Tax calculation will be finalised during checkout.
Sitarz R, Skierucha M, Mielko J, Offerhaus GJA, Maciejewski R, Polkowski WP. Gastric cancer: epidemiology, prevention, classification, and treatment. Cancer Manag Res. 2018;10:239–48.
Rugge M, Fassan M, Graham DY. Epidemiology of gastric cancer. In: Gastric cancer: Springer; 2015. p. 23–34.
Ajani JA, Lee J, Sano T, Janjigian YY, Fan D, Song S. Gastric adenocarcinoma. Nat Rev Dis Primers. 2017;3:17036.
Patel TN, Roy S, Ravi R. Gastric cancer and related epigenetic alterations. Ecancermedicalscience. 2017:11.
Takeshima H, Yamada H, Ushijima T. Cancer epigenetics: aberrant DNA methylation in cancer diagnosis and treatment. In: Oncogenomics: Elsevier; 2019. p. 65–76.
Kanwal R, Gupta K, Gupta S. Cancer epigenetics: an introduction. In: Cancer epigenetics: Springer; 2015. p. 3–25.
Li S, Zhu Y, Zhi L, Han X, Shen J, Liu Y, et al. DNA methylation variation trends during the embryonic development of chicken. PLoS One. 2016;11(7):e0159230.
Lim DH, Maher ER. DNA methylation: a form of epigenetic control of gene expression. Obstet Gynaecol. 2010;12(1):37–42.
Du J, Johnson LM, Jacobsen SE, Patel DJ. DNA methylation pathways and their crosstalk with histone methylation. Nat Rev Mol Cell Biol. 2015;16(9):519–32.
Tahara T, Arisawa T. DNA methylation as a molecular biomarker in gastric cancer. Epigenomics. 2015;7(3):475–86.
Li L, Ying J, Li H, Zhang Y, Shu X, Fan Y, et al. The human cadherin 11 is a pro-apoptotic tumor suppressor modulating cell stemness through Wnt/β-catenin signaling and silenced in common carcinomas. Oncogene. 2012;31(34):3901–12.
Carmona FJ, Villanueva A, Vidal A, Munoz C, Puertas S, Penin RM, et al. Epigenetic disruption of cadherin-11 in human cancer metastasis. J Pathol. 2012;228(2):230–40.
Yuan S, Li L, Xiang S, Jia H, Luo T. Cadherin-11 is inactivated due to promoter methylation and functions in colorectal cancer as a tumour suppressor. Cancer Manag Res. 2019;11:2517–29.
Lin YL, Gui SL, Ma JG. Aberrant methylation of CDH11 predicts a poor outcome for patients with bladder cancer. Oncol Lett. 2015;10(2):647–52.
Pasquale EB. Eph receptors and ephrins in cancer: bidirectional signalling and beyond. Nat Rev Cancer. 2010;10(3):165–80.
Chen X, Wang X, Wei X, Wang J. EphA5 protein, a potential marker for distinguishing histological grade and prognosis in ovarian serous carcinoma. J Ovarian Res. 2016;9(1):83.
Li S, Zhu Y, Ma C, Qiu Z, Zhang X, Kang Z, et al. Downregulation of EphA5 by promoter methylation in human prostate cancer. BMC Cancer. 2015;15(1):18.
Brantley-Sieders DM, Jiang A, Sarma K, Badu-Nkansah A, Walter DL, Shyr Y, et al. Eph/ephrin profiling in human breast cancer reveals significant associations between expression level and clinical outcome. PLoS One. 2011;6(9):e24426.
Hwang J-A, Kim Y, Hong S-H, Lee J, Cho YG, Han J-Y, et al. Epigenetic inactivation of heparan sulfate (glucosamine) 3-O-sulfotransferase 2 in lung cancer and its role in tumorigenesis. PLoS One. 2013;8(11):e79634.
Vijaya Kumar A, Salem Gassar E, Spillmann D, Stock C, Sen YP, Zhang T, et al. HS3ST2 modulates breast cancer cell invasiveness via MAP kinase-and Tcf4 (Tcf7l2)-dependent regulation of protease and cadherin expression. Int J Cancer. 2014;135(11):2579–92.
Miyamoto K, Asada K, Fukutomi T, Okochi E, Yagi Y, Hasegawa T, et al. Methylation-associated silencing of heparan sulfate D-glucosaminyl 3-O-sulfotransferase-2 (3-OST-2) in human breast, colon, lung and pancreatic cancers. Oncogene. 2003;22(2):274–80.
Lim EH, Ng SL, Li JL, Chang AR, Ng J, Ilancheran A, et al. Cervical dysplasia: assessing methylation status (Methylight) of CCNA1, DAPK1, HS3ST2, PAX1 and TFPI2 to improve diagnostic accuracy. Gynecol Oncol. 2010;119(2):225–31.
Leontiou CA, Hadjidaniel MD, Mina P, Antoniou P, Ioannides M, Patsalis PC. Bisulfite conversion of DNA: performance comparison of different kits and methylation quantitation of epigenetic biomarkers that have the potential to be used in non-invasive prenatal testing. PLoS One. 2015;10(8):e0135058.
Organization WH. Global cancer rates could increase by 50% to 15 million by 2020. In: Global cancer rates could increase by 50% to 15 million by 2020; 2003.
Ferreira HJ, Esteller M. CpG islands in cancer: heads, tails, and sides. In: CpG Islands: Springer; 2018. p. 49–80.
Wang X, Xu H, Wu Z, Chen X, Wang J. Decreased expression of EphA5 is associated with Fuhrman nuclear grade and pathological tumour stage in cc RCC. Int J Exp Pathol. 2017;98(1):34–9.
Sun B, Wu J, Zhang T, Wang C. High-resolution analysis of genomic profiles of hepatocellular carcinoma cells with differential osteopontin expression. Cancer Biol Ther. 2008;7(3):387–91.
Wu J-C, Sun B-S, Ren N, Ye Q-H, Qin L-X. Genomic aberrations in hepatocellular carcinoma related to osteopontin expression detected by array-CGH. J Cancer Res Clin Oncol. 2010;136(4):595–601.
This work was funded and supported by Liver and Gastrointestinal Diseases Research Center of Tabriz University of Medical Sciences (Grantnumber: 61877) and Molecular Medicine Research Center, Bio‐medicine Institute, Tabriz University of Medical Sciences, Tabriz, Iran.
The present study was approved by the Ethics and Research Committees of the Tabriz University of Medical Sciences.
Conflict of Interest
The authors report no conflicts of interest in this work.
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
About this article
Cite this article
Eyvazi, S., Khamaneh, A.M., Tarhriz, V. et al. CpG Islands Methylation Analysis of CDH11, EphA5, and HS3ST2 Genes in Gastric Adenocarcinoma Patients. J Gastrointest Canc 51, 579–583 (2020). https://doi.org/10.1007/s12029-019-00290-1
- Gastric adenocarcinoma
- Hyper methylation