Correlation between the promoter methylation status of ATP-binding cassette sub-family G member 2 and drug sensitivity in colorectal cancer cell lines

Moon,Hyun‑Hye; Kim,Sung‑Hee; Ku,Ja‑Lok

doi:10.3892/or.2015.4342

Correlation between the promoter methylation status of ATP-binding cassette sub-family G member 2 and drug sensitivity in colorectal cancer cell lines

Authors:
- Hyun‑Hye Moon
- Sung‑Hee Kim
- Ja‑Lok Ku
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Affiliations: Laboratory of Cell Biology, Cancer Research Institute, Seoul National University College of Medicine, Seoul 110‑799, Republic of Korea
Published online on: October 21, 2015 https://doi.org/10.3892/or.2015.4342
Pages: 298-306

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Abstract

Resistance to chemotherapeutic agents has been considered as a major reason for the high incidence rate of recurrence and metastasis suffered by colorectal cancer (CRC) patients. ATP‑binding cassette sub‑family G member 2 (ABCG2) is involved in drug resistance. DNA methylation of the ABCG2 promoter site has a significant influence on the regulation of epigenetic gene expression. In the present study, we investigated whether the methylation status of the ABCG2 promoter is related to drug sensitivity in CRC cell lines. In order to examine the ABCG2 expression level and identify the methylation status, RT‑PCR, qRT‑PCR analysis, MS‑PCR and bisulfite sequencing were conducted on 32 CRC cell lines. SNU‑C4, LS174T and NCI‑H716 were selected as low ABCG2‑expressing and high promoter methylated cell lines. The cell proliferation assay for 5‑fluorouracil, oxaliplatin and irinotecan was performed after 5‑aza‑2'‑deoxycytidine (5‑aza) treatment in these cell lines. In the 32 CRC cell lines, 25% of the cell lines expressed low or no ABCG2 expression. Of these cell lines, SNU‑C4, LS174T and NCI‑H716 were hypermethylated at the promoter region, ~20%. Demethylation of ABCG2 was induced by 5‑aza, which enhanced the ABCG2 expression level and influenced the cell proliferation similar to treatment with the anticancer agents. Our data suggest that the ABCG2 expression level regulated by methylation is related to anticancer drug sensitivity. Based on these results, it can be applied to predict the anticancer drug response.

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1	Jemal A, Bray F, Center MM, Ferlay J, Ward E and Forman D: Global cancer statistics. CA Cancer J Clin. 61:69–90. 2011. View Article : Google Scholar : PubMed/NCBI
2	André T, Boni C, Mounedji-Boudiaf L, Navarro M, Tabernero J, Hickish T, Topham C, Zaninelli M, Clingan P, Bridgewater J, et al Multicenter International Study of Oxaliplatin/5-Fluorouracil/Leucovorin in the Adjuvant Treatment of Colon Cancer (MOSAIC) Investigators: Oxaliplatin, fluorouracil and leucovorin as adjuvant treatment for colon cancer. N Engl J Med. 350:2343–2351. 2004. View Article : Google Scholar
3	Violette S, Poulain L, Dussaulx E, Pepin D, Faussat AM, Chambaz J, Lacorte JM, Staedel C and Lesuffleur T: Resistance of colon cancer cells to long-term 5-fluorouracil exposure is correlated to the relative level of Bcl-2 and Bcl-X(L) in addition to Bax and p53 status. Int J Cancer. 98:498–504. 2002. View Article : Google Scholar : PubMed/NCBI
4	Cazin JL, Gosselin P, Cappelaere P, Robert J and Demaille A: Drug resistance in oncology: From concepts to applications. J Cancer Res Clin Oncol. 119:76–86. 1992. View Article : Google Scholar : PubMed/NCBI
5	Gottesman MM: Mechanisms of cancer drug resistance. Annu Rev Med. 53:615–627. 2002. View Article : Google Scholar : PubMed/NCBI
6	Eijdems EW, De Haas M, Coco-Martin JM, Ottenheim CP, Zaman GJ, Dauwerse HG, Breuning MH, Twentyman PR, Borst P and Baas F: Mechanisms of MRP over-expression in four human lung-cancer cell lines and analysis of the MRP amplicon. Int J Cancer. 60:676–684. 1995. View Article : Google Scholar : PubMed/NCBI
7	Bailey-Dell KJ, Hassel B, Doyle LA and Ross DD: Promoter characterization and genomic organization of the human breast cancer resistance protein (ATP-binding cassette transporter G2) gene. Biochim Biophys Acta. 1520:234–241. 2001. View Article : Google Scholar : PubMed/NCBI
8	Litman T, Brangi M, Hudson E, Fetsch P, Abati A, Ross DD, Miyake K, Resau JH and Bates SE: The multidrug-resistant phenotype associated with overexpression of the new ABC half transporter, MXR (ABCG2). J Cell Sci. 113:2011–2021. 2000.
9	Xu J, Liu Y, Yang Y, Bates S and Zhang JT: Characterization of oligomeric human half-ABC transporter ATP-binding cassette G2. J Biol Chem. 279:19781–19789. 2004. View Article : Google Scholar : PubMed/NCBI
10	Turner JG, Gump JL, Zhang C, Cook JM, Marchion D, Hazlehurst L, Munster P, Schell MJ, Dalton WS and Sullivan DM: ABCG2 expression, function, and promoter methylation in human multiple myeloma. Blood. 108:3881–3889. 2006. View Article : Google Scholar : PubMed/NCBI
11	To KK, Zhan Z and Bates SE: Aberrant promoter methylation of the ABCG2 gene in renal carcinoma. Mol Cell Biol. 26:8572–8585. 2006. View Article : Google Scholar : PubMed/NCBI
12	To KK, Zhan Z, Litman T and Bates SE: Regulation of ABCG2 expression at the 3′ untranslated region of its mRNA through modulation of transcript stability and protein translation by a putative microRNA in the S1 colon cancer cell line. Mol Cell Biol. 28:5147–5161. 2008. View Article : Google Scholar : PubMed/NCBI
13	Nakano H, Nakamura Y, Soda H, Kamikatahira M, Uchida K, Takasu M, Kitazaki T, Yamaguchi H, Nakatomi K, Yanagihara K, et al: Methylation status of breast cancer resistance protein detected by methylation-specific polymerase chain reaction analysis is correlated inversely with its expression in drug-resistant lung cancer cells. Cancer. 112:1122–1130. 2008. View Article : Google Scholar : PubMed/NCBI
14	Mirza S, Sharma G, Pandya P and Ralhan R: Demethylating agent 5-aza-2-deoxycytidine enhances susceptibility of breast cancer cells to anticancer agents. Mol Cell Biochem. 342:101–109. 2010. View Article : Google Scholar : PubMed/NCBI
15	Doyle LA, Yang W, Abruzzo LV, Krogmann T, Gao Y, Rishi AK and Ross DD: A multidrug resistance transporter from human MCF-7 breast cancer cells. Proc Natl Acad Sci USA. 95:15665–15670. 1998. View Article : Google Scholar : PubMed/NCBI
16	Allen JD and Schinkel AH: Multidrug resistance and pharmacological protection mediated by the breast cancer resistance protein (BCRP/ABCG2). Mol Cancer Ther. 1:427–434. 2002.PubMed/NCBI
17	Yoh K, Ishii G, Yokose T, Minegishi Y, Tsuta K, Goto K, Nishiwaki Y, Kodama T, Suga M and Ochiai A: Breast cancer resistance protein impacts clinical outcome in platinum-based chemotherapy for advanced non-small cell lung cancer. Clin Cancer Res. 10:1691–1697. 2004. View Article : Google Scholar : PubMed/NCBI
18	König SK, Herzog M, Theile D, Zembruski N, Haefeli WE and Weiss J: Impact of drug transporters on cellular resistance towards saquinavir and darunavir. J Antimicrob Chemother. 65:2319–2328. 2010. View Article : Google Scholar : PubMed/NCBI
19	Langmann T, Mauerer R, Zahn A, Moehle C, Probst M, Stremmel W and Schmitz G: Real-time reverse transcription-PCR expression profiling of the complete human ATP-binding cassette transporter superfamily in various tissues. Clin Chem. 49:230–238. 2003. View Article : Google Scholar : PubMed/NCBI
20	Hendrich B and Bird A: Identification and characterization of a family of mammalian methyl-CpG binding proteins. Mol Cell Biol. 18:6538–6547. 1998. View Article : Google Scholar : PubMed/NCBI
21	Watt F and Molloy PL: Cytosine methylation prevents binding to DNA of a HeLa cell transcription factor required for optimal expression of the adenovirus major late promoter. Genes Dev. 2:1136–1143. 1988. View Article : Google Scholar : PubMed/NCBI
22	Perego P, De Cesare M, De Isabella P, Carenini N, Beggiolin G, Pezzoni G, Palumbo M, Tartaglia L, Pratesi G, Pisano C, et al: A novel 7-modified camptothecin analog overcomes breast cancer resistance protein-associated resistance in a mitoxantrone-selected colon carcinoma cell line. Cancer Res. 61:6034–6037. 2001.PubMed/NCBI
23	Boyer J, McLean EG, Aroori S, Wilson P, McCulla A, Carey PD, Longley DB and Johnston PG: Characterization of p53 wild-type and null isogenic colorectal cancer cell lines resistant to 5-fluorouracil, oxaliplatin and irinotecan. Clin Cancer Res. 10:2158–2167. 2004. View Article : Google Scholar : PubMed/NCBI
24	Yuan J, Lv H, Peng B, Wang C, Yu Y and He Z: Role of BCRP as a biomarker for predicting resistance to 5-fluorouracil in breast cancer. Cancer Chemother Pharmacol. 63:1103–1110. 2009. View Article : Google Scholar