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p16 Methylation is associated with chemosensitivity to fluorouracil in patients with advanced gastric cancer

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Abstract

No effective biomarkers have been confirmed to predict chemosensitivity for patients with gastric cancer. The purpose of this study was to investigate whether DNA methylation is associated with chemosensitivity in patients with gastric cancer. Tumors and matched non-tumor biopsy tissues collected from 134 advanced gastric cancer (AGC) patients prior to fluorouracil-based chemotherapy were retrospectively analyzed. The methylation status of p16, E-cadherin (CDH1), MGMT (O-6-methylguanine-DNA methyltransferase), and human mutL homolog 1 (hMLH1) was evaluated using a Methylight assay, and the association between p16 methylation and the sensitivity of 5-fluorouracil in cell lines was determined by in vitro assay. The methylation of p16 (17.9 vs. 0 %, P = 0.002), CDH1 (20.9 vs. 2.2 %, P < 0.001), MGMT (17.9 vs. 0 %, P = 0.052), and hMLH1 (14.9 vs. 2.2 %, P = 0.024) was more common in gastric cancer tissues (n = 134) than in non-tumor tissues (n = 46). For all patients, a reverse correlation was only found between p16 methylation and clinical response (P = 0.017), which suggested that p16 methylation might be associated with chemosensitivity of fluorouracil in gastric cancer patients. Results from in vitro experiments demonstrated that p16 methylation was closely correlated with the sensitivity of 5-fluorouracil in gastric cancer cells. The present results indicated that the methylation of p16, CDH1, MGMT, and hMLH1 was both frequent and specific in gastric cancer tissues. p16 Methylation might be used to predict chemosensitivity of fluorouracil for patients with AGC when validated in large samples in the future.

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Acknowledgments

This work was supported by National Natural Science Foundation of China (No. 81172110), National High Technology Research and Development Program (No. 2012AA 02A 504), and Beijing Municipal Science and Technology Commission Program (No. Z11110706730000).

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The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education), Department of GI Oncology, Peking University Cancer Hospital and Institute, No. 52, Fucheng Road, Haidian District, Beijing, 100142, China

    Mingming Wang, Yilin Li, Jing Gao, Yanyan Li & Lin Shen

  2. Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education), Division of Etiology, Peking University Cancer Hospital and Institute, No. 52, Fucheng Road, Haidian District, Beijing, 100142, China

    Jing Zhou, Liankun Gu & Dajun Deng

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  1. Mingming Wang
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Correspondence to Lin Shen.

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12032_2014_988_MOESM1_ESM.tif

PFS and OS curves for 134 patients treated with fluorouracil-based chemotherapy as indicated by the methylation status of (A) p16, (B) CDH1, (C) MGMT, and (D) hMLH1 genes1 (TIFF 2977 kb)

12032_2014_988_MOESM2_ESM.tif

Analysis of p16 gene methylation. (A) CpG regions used for p16 bisulfite genomic sequencing analysis (35 CpGs): vertical bars indicate dinucleotide CpGs, and solid lines indicate the probe sequences used for Methylight analysis (CpG 17–24). (B) Representative results of bisulfite genomic sequencing of p16 gene. Cloned PCR products from 5 non-tumor and 10 tumor tissue samples (positive = 5, negative = 5) were examined on the basis of the Methylight results. The PCR products were cloned into the pCR4-Topo vector (Invitrogen, Carlsbad, CA, U.S.), and twelve clones were randomly chosen and sequenced. Black bars denote methylated CpG sites. ATG, start codon2 (TIFF 31522 kb)

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Wang, M., Li, Y., Gao, J. et al. p16 Methylation is associated with chemosensitivity to fluorouracil in patients with advanced gastric cancer. Med Oncol 31, 988 (2014). https://doi.org/10.1007/s12032-014-0988-2

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