The aim of this prospective study was to evaluate the feasibility of predicting GC metastasis using CDH1, GFRA1, P16 and ZNF382 DNA methylation as biomarkers.
198 GC patients without metastasis at the time of surgery resection were recruited into the double-blind cohort (NCT02159339). Gene methylation was analysed using MethyLight assays. GC metastasis and survival data were obtained from 178 patients with 94.7% compliance during follow-up.
Twenty six cases of metastasis and 5 cases of recurrence were observed in 178 cases (17.4%) during the follow-up (median, 62.7 months). The GC metastasis rate for GFRA1 methylation-positive patients was significantly reduced compared with GFRA1 methylation-negative patients (odds ratio [OR]: 0.23, 95% confidence interval [CI] 0.08–0.66). Similar results were also observed using ZNF382 methylation as a predictor (OR: 0.17, 95% CI 0.06–0.47). A risk score including methylation of GFRA1 and ZNF382 was generated. The metastasis rate was significantly increased in high-risk GC patients (OR: 4.71, 95% CI: 1.85–12.00). GC patients with high risk had a shorter overall survival, especially for patients with stage I GC (P = 0.024).
The combination of GFRA1 and ZNF382 methylation is a biomarker panel for the prediction of GC metastasis.
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Ferlay, J., Soerjomataram, I., Dikshit, R., Eser, S., Mathers, C., Rebelo, M. et al. Cancer incidence and mortality worldwide: sources, methods and major patterns in GLOBOCAN 2012. Int. J. Cancer 136, E359–E386 (2015).
Bang, Y. J., Kim, Y. W., Yang, H. K., Chung, H. C., Park, Y. K., Lee, K. H. et al. Adjuvant capecitabine and oxaliplatin for gastric cancer after D2 gastrectomy (CLASSIC): a phase 3 open-label, randomised controlled trial. Lancet 379, 315–321 (2012).
Ajani, J. A., D'Amico, T. A., Almhanna, K., Bentrem, D. J., Chao, J., Das, P. et al. Gastric cancer, version 3.2016, NCCN clinical practice guidelines in oncology. J. Natl. Compr. Canc. Netw. 14, 1286–1312 (2016).
Aguirre-Ghiso, J. A., Bragado, P. & Sosa, M. S. Metastasis awakening: targeting dormant cancer. Nat. Med. 19, 276–277 (2013).
Polzer, B. & Klein, C. A. Metastasis awakening: the challenges of targeting minimal residual cancer. Nat. Med. 19, 274–275 (2013).
Bass, A. J. Comprehensive molecular characterization of gastric adenocarcinoma. Nature 513, 202–209 (2014).
Shimizu, T., Marusawa, H., Matsumoto, Y., Inuzuka, T., Ikeda, A., Fujii, Y. et al. Accumulation of somatic mutations in TP53 in gastric epithelium with Helicobacter pylori infection. Gastroenterology 147, 407–417 (2014).
Shigematsu, Y., Niwa, T., Yamashita, S., Taniguchi, H., Kushima, R., Katai, H. et al. Identification of a DNA methylation marker that detects the presence of lymph node metastases of gastric cancers. Onco. Lett. 4, 268–274 (2012).
Terashima, M., Ichikawa, W., Ochiai, A., Katada, K., Kurahashi, I., Sakuramoto, S. et al. TOP2A, GGH, PECAM1 are associated with hemotaogenous, lymph node, and peritoneal recurrence in stage II/III gastric cancer patients enrolled in the ACTS-GC study. Oncotarget. 8, 57574–57582 (2017).
Belinsky, S. A. Gene-promoter hypermethylation as a biomarker in lung cancer. Nat. Rev. Cancer 4, 707–717 (2004).
Liu, Z. J., Zhang, J., Gao, Y. H., Pei, L. R., Zhou, J., Gu, L. K. et al. Large-scale characterization of DNA methylation changes in human gastric carcinomas with and without metastasis. Clin. Cancer. Res. 20, 4598–4612 (2014).
Beroukhim, R., Mermel, C. H., Porter, D., Wei, G., Raychaudhuri, S., Donovan, J. et al. The landscape of somatic copy-number alteration across human cancers. Nature 463, 899–905 (2010).
Chen, S., Sanjana, N. E., Zheng, K., Shalem, O., Lee, K., Shi, X. et al. Genome-wide CRISPR screen in a mouse model of tumor growth and metastasis. Cell 160, 1246–1260 (2015).
Cui, C. H., Gan, Y., Gu, L. K., Wilson, J., Liu, Z. J., Zhang, B. Z. et al. P16-specific DNA methylation by engineered zinc finger methyltransferase inactivates gene transcription and promotes cancer metastasis. Genome. Biol. 16, 252 (2015).
Grady, W., Willis, J., Guilford, P., Dunbier, A., Toro, T., Lynch, H. et al. Methylation of the CDH1 promoter as the second genetic hit in hereditary diffuse gastric cancer. Nat. Genet. 26, 16–17 (2000).
Corso, G., Carvalho, J., Marrelli, D., Vindigni, C., Carvalho, B., Seruca, R. et al. Somatic mutations and deletions of the E-cadherin gene predict poor survival of patients with gastric cancer. J. Clin. Oncol. 31, 868–875 (2013).
Sobin, L., Gospodarowicz, M., Wittekind, C. (eds). TNM Classification of Malignant Tumours, 7th edn. International Union Against Cancer (UICC). (Wiley Press, New York, USA, 2009).
International Cancer Genome, C., Hudson, T. J., Anderson, W., Artez, A., Barker, A. D., Bell, C. et al. International network of cancer genome projects. Nature 464, 993–998 (2010).
Eads C. & Laird P. Combined bisulfite restriction analysis (COBRA). in DNA Methylation Protocols (Methods in Molecular Biology, (eds Mills K., Ramsahoye B.) vol 200, pp 53–70 (Humana Press, Totowa, New Jersey, USA, 2002).
Toyooka, K. O., Toyooka, S., Maitra, A., Feng, Q., Kiviat, N. C., Smith, A. et al. Establishment and validation of real-time polymerase chain reaction method for CDH1 promoter methylation. Am J Pathol 161, 629–634 (2002).
Zhou, J., Cao, J., Lu, Z. M., Liu, H. W. & Deng, D. J. A 115-bp MethyLight assay for detection ofp16 (CDKN2A) methylation as a diagnostic biomarker in human tissues. BMC Medical Genetics 12, 67 (2011).
Widschwendter, M., Siegmund, K. D., Müller, H. M., Fiegl, H., Marth, C., Muller-Holzner, E. et al. Association of breast cancer DNA methylation profiles with hormone receptor status and response to tamoxifen. Cancer Res. 64, 3807–3813 (2004).
Liu, Z., Zhou, J., Gu, L. & Deng, D. Significant impact of amount of PCR input templates on various PCR-based DNA methylation analysis and countermeasure. Oncotarget. 7, 56447–56455 (2016).
Donohue, M. C., Overholser, R., Xu, R. & Vaida, F. Conditional Akaike information under generalized linear and proportional hazards mixed models. Biometrika 98, 685–700 (2011).
Wang, Y. W., Zhu, M. L., Wang, R. F., Xue, W. J., Zhu, X. R., Wang, L. F. et al. Predictable factors for lymph node metastasis in early gastric cancer analysis of clinicopathologic factors and biological markers. Tumor Biol. 37, 8567–8578 (2016).
Motoyama, K., Inoue, H., Mimori, K., Tanaka, F., Kojima, K., Uetake, H. et al. Clinicopathological and prognostic significance of PDCD4 and microRNA-21 in human gastric cancer. Int. J. Oncol 36, 1089–1095 (2010).
Tanaka, M., Kitajima, Y., Edakuni, G., Sato, S. & Miyazaki, K. Abnormal expression of E-cadherin and beta-catenin may be a molecular marker of submucosal invasion and lymph node metastasis in early gastric cancer. Br. J. Surg. 89, 236–244 (2002).
Arigami, T., Natsugoe, S., Uenosono, Y., Yanagita, S., Arima, H., Hirata, M. et al. CCR7 and CXCR4 expression predicts lymph node status including micrometastasis in gastric cancer. Int. J. Oncol. 35, 19–24 (2009).
Shen, Z., Ye, Y., Dong, L., Vainionpää, S., Mustonen, H., Puolakkainen, P. et al. Kindlin-2: a novel adhesion protein related to tumor invasion, lymph node metastasis, and patient outcome in gastric cancer. Am. J. Surg. 203, 222–229 (2012).
Cheng, Y., Geng, H., Cheng, S. H., Liang, P., Bai, Y., Li, J. et al. KRAB zinc finger protein ZNF382 is a proapoptotic tumor suppressor that represses multiple oncogenes and is commonly silenced in multiple carcinomas. Cancer Res. 70, 6516–6526 (2010).
Airaksinen, M. S. & Saarma, M. The GDNF family: signalling, biological functions and therapeutic value. Nat. Rev. Neurosci. 3, 383–394 (2002).
Esseghir, S., Todd, S. K., Hunt, T., Poulsom, R., Plaza-Menacho, I., Reis-Filho, J. S. et al. A role for glial cell derived neurotrophic factor induced expression by inflammatory cytokines and RET/GFR alpha1 receptor up-regulation in breast cancer. Cancer Res. 67, 11732–11741 (2007).
Cavel, O., Shomron, O., Shabtay, A., Vital, J., Trejo-Leider, L., Weizman, N. et al. Endoneurial macrophages induce perineural invasion of pancreatic cancer cells by secretion of GDNF and activation of RET tyrosine kinase receptor. Cancer Res. 72, 5733–5743 (2012).
The authors declare no competing interests.
Ethics approval and consent to participate
The Institution Review Board of Peking University Cancer Hospital & Institute approved this study and was carried out in accordance with the principles outlined in the Declaration of Helsinki. Informed consent was obtained from each patient prior to their inclusion in the study.
This work is supported by grants from the National Natural Science Foundation of China (no. 81402031), the Beijing Municipal and Technology Commission (Z151100001615022), the Science Foundation of Peking University Cancer Hospital (2017–25) and the Beijing Municipal Commission of Health and Family Planning (PXM2018_026279_000005).
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