Alteration of gene expression and DNA methylation in drug-resistant gastric cancer

Maeda,Osamu; Ando,Takafumi; Ohmiya,Naoki; Ishiguro,Kazuhiro; Watanabe,Osamu; Miyahara,Ryoji; Hibi,Yoko; Nagai,Taku; Yamada,Kiyofumi; Goto,Hidemi

doi:10.3892/or.2014.3014

Alteration of gene expression and DNA methylation in drug-resistant gastric cancer

Authors:
- Osamu Maeda
- Takafumi Ando
- Naoki Ohmiya
- Kazuhiro Ishiguro
- Osamu Watanabe
- Ryoji Miyahara
- Yoko Hibi
- Taku Nagai
- Kiyofumi Yamada
- Hidemi Goto
View Affiliations

Affiliations: Department of Advanced Research of Gastroenterology, Nagoya University Graduate School of Medicine, Showa-ku, Nagoya, Japan, Department of Gastroenterology and Hepatology, Nagoya University Graduate School of Medicine, Showa-ku, Nagoya, Japan, Department of Gastroenterology, School of Medicine, Fujita Health University, Toyoake, Aichi, Japan, Department of Hospital Pharmacy, Nagoya University Hospital, Nagoya, Japan
Published online on: February 6, 2014 https://doi.org/10.3892/or.2014.3014
Pages: 1883-1890

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

The mechanisms of drug resistance in cancer are not fully elucidated. To study the drug resistance of gastric cancer, we analyzed gene expression and DNA methylation profiles of 5-fluorouracil (5-FU)- and cisplatin (CDDP)-resistant gastric cancer cells and biopsy specimens. Drug-resistant gastric cancer cells were established with culture for >10 months in a medium containing 5-FU or CDDP. Endoscopic biopsy specimens were obtained from gastric cancer patients who underwent chemotherapy with oral fluoropyrimidine S-1 and CDDP. Gene expression and DNA methylation analyses were performed using microarray, and validated using real-time PCR and pyrosequencing, respectively. Out of 17,933 genes, 541 genes commonly increased and 569 genes decreased in both 5-FU- and CDDP-resistant AGS cells. Genes with expression changed by drugs were related to GO term ‘extracellular region’ and ‘p53 signaling pathway’ in both 5-FU- and CDDP-treated cells. Expression of 15 genes including KLK13 increased and 12 genes including ETV7 decreased, in both drug-resistant cells and biopsy specimens of two patients after chemotherapy. Out of 10,365 genes evaluated with both expression microarray and methylation microarray, 74 genes were hypermethylated and downregulated, or hypomethylated and upregulated in either 5-FU-resistant or CDDP-resistant cells. Of these genes, expression of 21 genes including FSCN1, CPT1C and NOTCH3, increased from treatment with a demethylating agent. There are alterations of gene expression and DNA methylation in drug-resistant gastric cancer; they may be related to mechanisms of drug resistance and may be useful as biomarkers of gastric cancer drug sensitivity.

View Figures

View References

1	Ferlay J, Shin HR, Bray F, Forman D, Mathers C and Parkin DM: Estimates of worldwide burden of cancer in 2008: GLOBOCAN 2008. Int J Cancer. 127:2893–2917. 2010. View Article : Google Scholar : PubMed/NCBI
2	Okines A, Verheij M, Allum W, Cunningham D and Cervantes A; ESMO Guidelines Working Group. Gastric cancer: ESMO Clinical Practice Guidelines for diagnosis, treatment and follow-up. Ann Oncol. 21(Suppl 5): v50–v54. 2010. View Article : Google Scholar : PubMed/NCBI
3	Zhang D and Fan D: New insights into the mechanisms of gastric cancer multidrug resistance and future perspectives. Future Oncol. 6:527–537. 2010. View Article : Google Scholar : PubMed/NCBI
4	Taby R and Issa JP: Cancer epigenetics. CA Cancer J Clin. 60:376–392. 2010. View Article : Google Scholar : PubMed/NCBI
5	Ushijima T: Epigenetic field for cancerization. J Biochem Mol Biol. 40:142–150. 2007. View Article : Google Scholar
6	Shen L, Kondo Y, Ahmed S, et al: Drug sensitivity prediction by CpG island methylation profile in the NCI-60 cancer cell line panel. Cancer Res. 67:11335–11343. 2007. View Article : Google Scholar : PubMed/NCBI
7	Yamashita S, Takahashi S, McDonell N, et al: Methylation silencing of transforming growth factor-β receptor type II in rat prostate cancers. Cancer Res. 68:2112–2121. 2008.
8	Koizumi W, Narahara H, Hara T, et al: S-1 plus cisplatin versus S-1 alone for first-line treatment of advanced gastric cancer (SPIRITS trial): a phase III trial. Lancet Oncol. 9:215–221. 2008. View Article : Google Scholar : PubMed/NCBI
9	Bibikova M, Barnes B, Tsan C, et al: High density DNA methylation array with single CpG site resolution. Genomics. 98:288–295. 2011. View Article : Google Scholar : PubMed/NCBI
10	Colella S, Shen L, Baggerly KA, Issa JP and Krahe R: Sensitive and quantitative universal Pyrosequencing methylation analysis of CpG sites. Biotechniques. 35:146–150. 2003.PubMed/NCBI
11	An B, Kondo Y, Okamoto Y, et al: Characteristic methylation profile in CpG island methylator phenotype-negative distal colorectal cancers. Int J Cancer. 127:2095–2105. 2010. View Article : Google Scholar : PubMed/NCBI
12	Shen L, Guo Y, Chen X, Ahmed S and Issa JP: Optimizing annealing temperature overcomes bias in bisulfite PCR methylation analysis. Biotechniques. 42:48–58. 2007. View Article : Google Scholar : PubMed/NCBI
13	Chou RH, Lin SC, Wen HC, Wu CW and Chang WS: Epigenetic activation of human kallikrein 13 enhances malignancy of lung adenocarcinoma by promoting N-cadherin expression and laminin degradation. Biochem Biophys Res Commun. 409:442–447. 2011. View Article : Google Scholar : PubMed/NCBI
14	Florou D, Mavridis K and Scorilas A: The kallikrein-related peptidase 13 (KLK13) gene is substantially up-regulated after exposure of gastric cancer cells to antineoplastic agents. Tumour Biol. 33:2069–2078. 2012.
15	Gu X, Shin BH, Akbarali Y, et al: Tel-2 is a novel transcriptional repressor related to the Ets factor Tel/ETV-6. J Biol Chem. 276:9421–9436. 2001. View Article : Google Scholar : PubMed/NCBI
16	Hanker LC, Karn T, Holtrich U, et al: Prognostic impact of fascin-1 (FSCN1) in epithelial ovarian cancer. Anticancer Res. 33:371–377. 2013.PubMed/NCBI
17	Tsai WC, Jin JS, Chang WK, et al: Association of cortactin and fascin-1 expression in gastric adenocarcinoma: correlation with clinicopathological parameters. J Histochem Cytochem. 55:955–962. 2007. View Article : Google Scholar : PubMed/NCBI
18	Reilly PT and Mak TW: Molecular pathways: tumor cells co-opt the brain-specific metabolism gene CPT1C to promote survival. Clin Cancer Res. 18:5850–5855. 2012. View Article : Google Scholar : PubMed/NCBI
19	Liu J, Fan H, Ma Y, et al: Notch1 is a 5-fluorouracil resistant and poor survival marker in human esophagus squamous cell carcinomas. PLoS One. 8:e561412013. View Article : Google Scholar : PubMed/NCBI
20	Ma XB, Jia XS, Liu YL, et al: Expression and role of Notch signalling in the regeneration of rat tracheal epithelium. Cell Prolif. 42:15–28. 2009. View Article : Google Scholar : PubMed/NCBI
21	Man CH, Wei-Man Lun S, Wai-Ying Hui J, et al: Inhibition of NOTCH3 signalling significantly enhances sensitivity to cisplatin in EBV-associated nasopharyngeal carcinoma. J Pathol. 226:471–481. 2012. View Article : Google Scholar : PubMed/NCBI
22	Rahman MT, Nakayama K, Rahman M, et al: Notch3 overexpression as potential therapeutic target in advanced stage chemoresistant ovarian cancer. Am J Clin Pathol. 138:535–544. 2012. View Article : Google Scholar : PubMed/NCBI