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Epigenetic alteration of p16 and retinoic acid receptor beta genes in the development of epithelial ovarian carcinoma

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Tumor Biology

Abstract

Silencing of tumor suppressor and tumor-related genes by promoter hypermethylation is one of the major events in ovarian carcinogenesis. In this study, we analyzed aberrant promoter methylation of p16 and RAR-β genes in 134 epithelial ovarian carcinomas (EOCs), 23 low malignant potential (LMP) tumors, 26 benign cystadenomas, and 15 normal ovarian tissues. Methylation was investigated by methylation-specific PCR (MSP), and the results were confirmed by bisulfite DNA sequencing. Relative gene expression of p16 and RAR-β was done using quantitative reverse transcriptase PCR (qRT-PCR) on 51 EOC cases, 9 LMP tumors, and 7 benign cystadenomas with 5 normal ovarian tissues. Aberrant methylation for p16 and RAR-β was present in 43 % (58/134) and 31 % (41/134) in carcinoma cases, 22 % (05/23) and 52 % (12/23) in LMP tumors, and 42 % (11/26) and 69 % (18/26) in benign cystadenomas. No methylation was observed in any of the normal ovarian tissues. The mRNA expression level of p16 and RAR-β was significantly downregulated in EOC and LMP tumors than the corresponding normal tissues whereas the expression level was normal in benign cystadenomas for p16 and slightly reduced for RAR-β. A significant correlation of p16 promoter methylation was observed with reduced gene expression in EOC. For RAR-β, no significant correlation was observed between promoter methylation and gene expression. Our results suggest that epigenetic alterations of p16 and RAR-β have an important role in ovarian carcinogenesis and that mechanism along with methylation plays a significant role in downregulation of RAR-β gene in ovarian cancer.

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Acknowledgments

The research work is supported by Department of Biotechnology, India, Grant No BT/PR 13440/MED/30/267/2009. We are grateful to Dr V Shanmugam (Research Assistant, NIMHANS) for helping with the statistical analysis.

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None.

Ethical clearance

The study was approved by Institutional Ethics Committee and all samples were collected after obtaining written, informed consent from patients.

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Authors and Affiliations

  1. Department of Biochemistry, Kidwai Memorial Institute of Oncology, Dr. M.H. Marigowda Road, Bangalore, 560029, India

    Rahul Bhagat, Sandeep Sriram Kumar, Shilpa Vaderhobli & Gawari Ramesh

  2. Department of Pathology, Kidwai Memorial Institute of Oncology, Dr. M.H. Marigowda Road, Bangalore, 560029, India

    Chennagiri S. Premalata

  3. Department of Gynaec Oncology, Kidwai Memorial Institute of Oncology, Dr. M.H. Marigowda Road, Bangalore, 560029, India

    Venkateshaiah Reddihalli Pallavi

  4. Department of Laboratory Medicine, Sri Shankara Cancer Hospital and Research Centre, Shankara Math Premises, Basavanagudi, Bangalore, 560004, India

    Lakshmi Krishnamoorthy

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  1. Rahul Bhagat
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Correspondence to Lakshmi Krishnamoorthy.

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Bhagat, R., Kumar, S.S., Vaderhobli, S. et al. Epigenetic alteration of p16 and retinoic acid receptor beta genes in the development of epithelial ovarian carcinoma. Tumor Biol. 35, 9069–9078 (2014). https://doi.org/10.1007/s13277-014-2136-1

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