Open Access Research article

High-throughput detection of aberrant imprint methylation in the ovarian cancer by the bisulphite PCR-Luminex method

Hitoshi Hiura1, Hiroaki Okae1, Hisato Kobayash2, Naoko Miyauchi1, Fumi Sato1, Akiko Sato3, Fumihiko Suzuki3, Satoru Nagase3, Junichi Sugawara3, Kunihiko Nakai4, Nobuo Yaegashi3 and Takahiro Arima1*

Author Affiliations

1 Department of Informative Genetics, Environment and Genome Research Center, Tohoku University Graduate School of Medicine, 2-1 Seiryo-cho, Aoba-ku, Sendai 980-8575, Japan

2 Department of BioScience, Tokyo University of Agriculture, 1-1-1 Sakuragaoka, Setagaya-ku, Tokyo 156-8502, Japan

3 Departments of Obstetrics and Gynecology, Tohoku University Graduate, School of Medicine, Sendai, Japan

4 Department of Development and Environmental Medicine, Tohoku University Graduate School of Medicine, Sendai, Japan

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BMC Medical Genomics 2012, 5:8  doi:10.1186/1755-8794-5-8

Published: 26 March 2012

Additional files

Additional file 1:

Table S1 The list of LOH, LOI and MOI in HOC. LOH, LOI and MOI determined using RFLP analysis of 8 imprinted genes are summarized. NC: Normal cells (NC1-4). NT: Normal ovarian tissues (NT1-7). CC: Cancer cell lines (CC1-21). CT: Cancer tissue (CT1-74).

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Additional file 2:

Figure S1 Validation of BPL analyses by comparison with COBRA assay. Examination of the imprinted DMRs by bisulphite PCR Luminex (BPL) and combined bisulphite PCR restriction analysis (COBRA) assay in DNA samples of ovarian cancer cell lines and normal cells. BPL: y-axis, COBRA: x-axis. The number was calculated by Spearman's rank method. GTL2 (C), ZDBF2 (D), LIT1 (E), ZAC (F), PEG3 (G) and SNRPN (H).

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Additional file 3:

Table S2 Sequences of PCR primers and restriction enzymes used for PCR-RFLP analysis.

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Additional file 4:

Table S3 Bisulphite PCR-Luminex and COBRA methylation profiles of the eight imprinted DMRs in the DNA of human ovarian cancer cells and normal ovarian tissues. Numbers in blue and black boldface indicate LOI and MOI, respectively. Luminex values indicate average methylation values at the sites tested. NC: Normal cells (OSE1, OSE2, OSE3, OSE4). CC: Cancer cell lines (CC1: OVCAR3, CC2: OMC3, CC3: CAOV3, CC4: JHOS2, CC5: HTOA, CC6: TOV112D, CC7: ES2, CC8: JHOC5, CC9: SKOV3, CC10: TOV21G, CC11: OV90, CC12: JHOS3, CC13: JHOS4, CC14: JHOC7, CC15: JHOC8, CC16: KF, CC17: KM, CC18: HAC, CC19: RMG, CC20: MCAS, CC21: MH. NT: Normal ovarian tissues (NT1-NT7). N: Normal, S: Serous adenocarcinoma, M: Mucinous adenocarcinoma, E: Endometrioid carcinoma, C: Clear cell carcinoma.

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