Aberrant methylation of microRNA-34b/c is a predictive marker of metachronous gastric cancer risk

Ryo Suzuki, Eiichiro Yamamoto, Masanori Nojima, Reo Maruyama, Hiro-O Yamano, Kenjiro Yoshikawa, Tomoaki Kimura, Taku Harada, Masami Ashida, Takeshi Niinuma, Akiko Sato, Katsuhiko Nosho, Hiroyuki Yamamoto, Masahiro Kai, Tamotsu Sugai, Kohzoh Imai, Hiromu Suzuki, Yasuhisa Shinomura, Ryo Suzuki, Eiichiro Yamamoto, Masanori Nojima, Reo Maruyama, Hiro-O Yamano, Kenjiro Yoshikawa, Tomoaki Kimura, Taku Harada, Masami Ashida, Takeshi Niinuma, Akiko Sato, Katsuhiko Nosho, Hiroyuki Yamamoto, Masahiro Kai, Tamotsu Sugai, Kohzoh Imai, Hiromu Suzuki, Yasuhisa Shinomura

Abstract

Background: Metachronous gastric cancer (GC) can develop after endoscopic resection of GC and cannot be predicted based on clinical signature. Aberrant DNA methylation in noncancerous gastric mucosa is strongly implicated in gastric carcinogenesis and could be a useful biomarker of GC risk. We evaluated the clinical utility of DNA methylation as a biomarker of metachronous GC risk.

Method: We carried out scheduled follow-up endoscopy in 129 patients after curative endoscopic resection of GC. Biopsy specimens were collected from noncancerous mucosa in the gastric antrum and body, after which quantitative methylation analysis of miR-34b/c, SFRP1, SFRP2, SFRP5, DKK2 and DKK3 was carried out using bisulfite pyrosequencing. The utility of the methylation for predicting the risk of metachronous GC development was assessed using Kaplan-Meier and Cox proportional hazards model analyses.

Results: During the follow-up period, 17 patients (13%) developed metachronous GCs. The cumulative incidence of metachronous GC was significantly higher among patients with elevated miR-34b/c, SFRP2 and DKK2 methylation in their gastric body. MiR-34b/c showed the strongest association with the risk of metachronous GC, and the cumulative incidence of metachronous GC was much higher in the high-miR-34b/c-methylation group than the low-methylation group. Multivariate analysis adjusted for age, sex, H. pylori status and pathological findings showed miR-34b/c methylation in gastric body to be an independent predictor of metachronous GC risk.

Conclusion: Our results suggest that methylation of miR-34b/c in the mucosa of the noncancerous gastric body may be a useful biomarker for predicting the risk of metachronous GC.

Figures

Fig. 1
Fig. 1
Profiles of participants in this study
Fig. 2
Fig. 2
Associations between DNA methylation and metachronous GC risk. aForest plot showing hazard ratios (closed circles) for developing metachronous GC and 95 % confidence intervals (bar lines). Univariate Cox proportional hazards model analysis was performed to assess the correlations between methylation of the six indicated genes and the incidence of metachronous GC. b Kaplan–Meier analysis of the effect of miR-34b/c methylation in the gastric body on metachronous GC-free survival (n = 129)
Fig. 3
Fig. 3
Association between miR-34b/c methylation and metachronous GC risk in H. pylori-positive and -negative patients. a Kaplan–Meier analysis of the effect of miR-34b/c methylation on metachronous GC-free survival among H. pylori-positive patients who underwent successful eradication after endoscopic treatment of their initial GC (n = 49). b Kaplan–Meier analysis of the effect of miR-34b/c methylation on metachronous GC-free survival among H. pylori-negative patients (n = 26)

References

    1. Ono H, Kondo H, Gotoda T, Shirao K, Yamaguchi H, Saito D, et al. Endoscopic mucosal resection for treatment of early gastric cancer. Gut. 2001;48:225–229. doi: 10.1136/gut.48.2.225.
    1. Soetikno R, Kaltenbach T, Yeh R, Gotoda T. Endoscopic mucosal resection for early cancers of the upper gastrointestinal tract. J Clin Oncol. 2005;23:4490–4498. doi: 10.1200/JCO.2005.19.935.
    1. Oka S, Tanaka S, Kaneko I, Mouri R, Hirata M, Kawamura T, et al. Advantage of endoscopic submucosal dissection compared with EMR for early gastric cancer. Gastrointest Endosc. 2006;64:877–883. doi: 10.1016/j.gie.2006.03.932.
    1. Isomoto H, Shikuwa S, Yamaguchi N, Fukuda E, Ikeda K, Nishiyama H, et al. Endoscopic submucosal dissection for early gastric cancer: a large-scale feasibility study. Gut. 2009;58:331–336. doi: 10.1136/gut.2008.165381.
    1. Nakajima T, Oda I, Gotoda T, Hamanaka H, Eguchi T, Yokoi C, et al. Metachronous gastric cancers after endoscopic resection: how effective is annual endoscopic surveillance? Gastric Cancer. 2006;9:93–98. doi: 10.1007/s10120-006-0372-9.
    1. Maehata Y, Nakamura S, Fujisawa K, Esaki M, Moriyama T, Asano K, et al. Long-term effect of Helicobacter pylori eradication on the development of metachronous gastric cancer after endoscopic resection of early gastric cancer. Gastrointest Endosc. 2012;75:39–46. doi: 10.1016/j.gie.2011.08.030.
    1. Kato M, Nishida T, Yamamoto K, Hayashi S, Kitamura S, Yabuta T, et al. Scheduled endoscopic surveillance controls secondary cancer after curative endoscopic resection for early gastric cancer: a multicentre retrospective cohort study by Osaka University ESD study group. Gut. 2012.
    1. Uemura N, Okamoto S, Yamamoto S, Matsumura N, Yamaguchi S, Yamakido M, et al. Helicobacter pylori infection and the development of gastric cancer. N Engl J Med. 2001;345:784–789. doi: 10.1056/NEJMoa001999.
    1. Fukase K, Kato M, Kikuchi S, Inoue K, Uemura N, Okamoto S, et al. Effect of eradication of Helicobacter pylori on incidence of metachronous gastric carcinoma after endoscopic resection of early gastric cancer: an open-label, randomised controlled trial. Lancet. 2008;372:392–397. doi: 10.1016/S0140-6736(08)61159-9.
    1. Suzuki H, Tokino T, Shinomura Y, Imai K, Toyota M. DNA methylation and cancer pathways in gastrointestinal tumors. Pharmacogenomics. 2008;9:1917–1928. doi: 10.2217/14622416.9.12.1917.
    1. Yamamoto E, Suzuki H, Takamaru H, Yamamoto H, Toyota M, Shinomura Y. Role of DNA methylation in the development of diffuse-type gastric cancer. Digestion. 2011;83:241–249. doi: 10.1159/000320453.
    1. Jones P, Baylin S. The epigenomics of cancer. Cell. 2007;128:683–692. doi: 10.1016/j.cell.2007.01.029.
    1. Maekita T, Nakazawa K, Mihara M, Nakajima T, Yanaoka K, Iguchi M, et al. High levels of aberrant DNA methylation in Helicobacter pylori-infected gastric mucosae and its possible association with gastric cancer risk. Clin Cancer Res. 2006;12:989–995. doi: 10.1158/1078-0432.CCR-05-2096.
    1. Yamamoto E, Toyota M, Suzuki H, Kondo Y, Sanomura T, Murayama Y, et al. LINE-1 hypomethylation is associated with increased CpG island methylation in Helicobacter pylori-related enlarged-fold gastritis. Cancer Epidemiol Biomarkers Prev. 2008;17:2555–2564. doi: 10.1158/1055-9965.EPI-08-0112.
    1. Nakajima T, Maekita T, Oda I, Gotoda T, Yamamoto S, Umemura S, et al. Higher methylation levels in gastric mucosae significantly correlate with higher risk of gastric cancers. Cancer Epidemiol Biomarkers Prev. 2006;15:2317–2321. doi: 10.1158/1055-9965.EPI-06-0436.
    1. Ushijima T, Hattori N. Molecular pathways: involvement of Helicobacter pylori-triggered inflammation in the formation of an epigenetic field defect, and its usefulness as cancer risk and exposure markers. Clin Cancer Res. 2012;18:923–929. doi: 10.1158/1078-0432.CCR-11-2011.
    1. Suzuki H, Yamamoto E, Nojima M, Kai M, Yamano H, Yoshikawa K, et al. Methylation-associated silencing of microRNA-34b/c in gastric cancer and its involvement in an epigenetic field defect. Carcinogenesis. 2010;31:2066–2073. doi: 10.1093/carcin/bgq203.
    1. Dixon MF, Genta RM, Yardley JH, Correa P. Classification and grading of gastritis. The updated Sydney system. In: International workshop on the histopathology of gastritis, Houston 1994. Am J Surg Pathol. 1996;20:1161.
    1. Toyota M, Suzuki H, Sasaki Y, Maruyama R, Imai K, Shinomura Y, et al. Epigenetic silencing of microRNA-34b/c and B-cell translocation gene 4 is associated with CpG island methylation in colorectal cancer. Cancer Res. 2008;68:4123–4132. doi: 10.1158/0008-5472.CAN-08-0325.
    1. Nojima M, Suzuki H, Toyota M, Watanabe Y, Maruyama R, Sasaki S, et al. Frequent epigenetic inactivation of SFRP genes and constitutive activation of wnt signaling in gastric cancer. Oncogene. 2007;26:4699–4713. doi: 10.1038/sj.onc.1210259.
    1. Sato H, Suzuki H, Toyota M, Nojima M, Maruyama R, Sasaki S, et al. Frequent epigenetic inactivation of DICKKOPF family genes in human gastrointestinal tumors. Carcinogenesis. 2007;28:2459–2466. doi: 10.1093/carcin/bgm178.
    1. Chan AO, Lam S, Wong BC, Yuen M, Yeung Y, Hui W, et al. Promoter methylation of E-cadherin gene in gastric mucosa associated with Helicobacter pylori infection and in gastric cancer. Gut. 2003;52:502–506. doi: 10.1136/gut.52.4.502.
    1. Cheng ASL, Li M, Kang W, Cheng V, Chou J, Lau S, et al. Helicobacter pylori causes epigenetic dysregulation of FOXD3 to promote gastric carcinogenesis. Gastroenterology. 2013;144:122, 133.e9.
    1. Hayashi Y, Tsujii M, Wang J, Kondo J, Akasaka T, Jin Y, et al. CagA mediates epigenetic regulation to attenuate let-7 expression in Helicobacter pylori-related carcinogenesis. Gut. 2012.
    1. Ando T, Yoshida T, Enomoto S, Asada K, Tatematsu M, Ichinose M, et al. DNA methylation of microRNA genes in gastric mucosae of gastric cancer patients: its possible involvement in the formation of epigenetic field defect. Int J Cancer. 2009;124:2367–2374. doi: 10.1002/ijc.24219.
    1. Nanjo S, Asada K, Yamashita S, Nakajima T, Nakazawa K, Maekita T, et al. Identification of gastric cancer risk markers that are informative in individuals with past H. pylori infection. Gastric Cancer. 2012;15:382–388. doi: 10.1007/s10120-011-0126-1.
    1. Takamaru H, Yamamoto E, Suzuki H, Nojima M, Maruyama R, Yamano HO, et al. Aberrant methylation of RASGRF1 is associated with an epigenetic field defect and increased risk of gastric cancer. Cancer Prev Res (Phila) 2012;5:1203–1212. doi: 10.1158/1940-6207.CAPR-12-0056.
    1. Suzuki H, Maruyama R, Yamamoto E, Kai M. DNA methylation and microRNA dysregulation in cancer. Mol Oncol. 2012;6:567–578. doi: 10.1016/j.molonc.2012.07.007.
    1. Niwa T, Tsukamoto T, Toyoda T, Mori A, Tanaka H, Maekita T, et al. Inflammatory processes triggered by Helicobacter pylori infection cause aberrant DNA methylation in gastric epithelial cells. Cancer Res. 2010;70:1430–1440. doi: 10.1158/0008-5472.CAN-09-2755.
    1. Chiba T, Marusawa H, Ushijima T. Inflammation-associated cancer development in digestive organs: mechanisms and roles for genetic and epigenetic modulation. Gastroenterology. 2012;143:550–563. doi: 10.1053/j.gastro.2012.07.009.
    1. Ikeno T, Ota H, Sugiyama A, Ishida K, Katsuyama T, Genta RM, et al. Helicobacter pylori-induced chronic active gastritis, intestinal metaplasia, and gastric ulcer in Mongolian gerbils. Am J Pathol. 1999;154:951–960. doi: 10.1016/S0002-9440(10)65343-6.
    1. Naylor GM, Gotoda T, Dixon M, Shimoda T, Gatta L, Owen R, et al. Why does Japan have a high incidence of gastric cancer? Comparison of gastritis between UK and Japanese patients. Gut. 2006;55:1545–1552. doi: 10.1136/gut.2005.080358.
    1. Wong B, Lam S, Wong W, Chen J, Zheng T, Feng R, et al. Helicobacter pylori eradication to prevent gastric cancer in a high-risk region of China: a randomized controlled trial. JAMA (Chicago, IL) 2004;291:187–194.
    1. Wu CY, Kuo KN, Wu MS, Chen YJ, Wang CB, Lin JT. Early Helicobacter pylori eradication decreases risk of gastric cancer in patients with peptic ulcer disease. Gastroenterology. 2009;137:1641, 8.e1–2.
    1. Chan AO, Peng JZ, Lam SK, Lai KC, Yuen MF, Cheung HK, et al. Eradication of Helicobacter pylori infection reverses E-cadherin promoter hypermethylation. Gut. 2006;55:463–468. doi: 10.1136/gut.2005.077776.
    1. Nakajima T, Enomoto S, Yamashita S, Ando T, Nakanishi Y, Nakazawa K, et al. Persistence of a component of DNA methylation in gastric mucosae after Helicobacter pylori eradication. J Gastroenterol. 2010;45:37–44. doi: 10.1007/s00535-009-0142-7.
    1. Perri F, Cotugno R, Piepoli A, Merla A, Quitadamo M, Gentile A, et al. Aberrant DNA methylation in non-neoplastic gastric mucosa of H. pylori infected patients and effect of eradication. Am J Gastroenterol. 2007;102:1361–1371. doi: 10.1111/j.1572-0241.2007.01284.x.
    1. Sepulveda A, Yao Y, Yan W, Park D, Kim J, Gooding W, et al. CpG methylation and reduced expression of O6-methylguanine DNA methyltransferase is associated with Helicobacter pylori infection. Gastroenterology. 2010;138:1836–1844. doi: 10.1053/j.gastro.2009.12.042.

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