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Molecular pathogenesis of human prolactinomas identified by gene expression profiling, RT-qPCR, and proteomic analyses

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Abstract

The molecular pathogenesis of prolactinomas has resisted elucidation; with the exception of a RAS mutation in a single aggressive prolactinoma, no mutational changes have been identified. In prolactinomas, a further obstacle has been the paucity of surgical specimens suitable for molecular analysis since prolactionomas are infrequently removed due to the availability and effectiveness of medical therapy. In the absence of mutational events, gene expression changes have been sought and detected. Using high-throughput analysis from a large bank of human pituitary adenomas, we examined these tumors according to their molecular profiles rather than traditional immunohistochemistry. We examined six prolactinomas and eight normal pituitary glands using oligonucleotide GeneChip microarrays, reverse transcription-real time quantitative polymerase chain reaction using 10 prolactinomas, and proteomic analysis to examine protein expression in four prolactinomas. Microarray analyses identified 726 unique genes that were statistically significantly different between prolactinomas and normal glands, whereas proteomic analysis identified four differently up-regulated and 19 down-regulated proteins. Several components of the Notch pathway were altered in prolactinomas, and there was an increased expression of the Pit-1 transcription factor, and the survival factor BAG1 but decreased E-cadherin and N-cadherin expression. Taken together, expression profiling and proteomic analyses have identified molecular features unique to prolactinomas that may contribute to their pathogenesis. In the current era of molecular medicine, these findings greatly enhance our understanding and supercede immunohistochemical diagnosis.

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Acknowledgements

The authors gratefully acknowledge financial assistance to Nelson M. Oyesiku, M.D., Ph.D., FACS, from the Department of Neurosurgery, to Carlos S. Moreno, PhD., from the National Institutes of Health (K22-CA96560 and R01-CA106826), to Paula M. Vertino, PhD from the National Institutes of Health (2RO1-CA077337), Michael, T. McCabe, PhD from the Frederick Gardner Cottrell Fellowship Program and to Dominic M. Desiderio, PhD., from the National Institutes of Heath (NS 42843), (RR-10522), (RR-14593) and NSF (DBI 9604633). The authors thank the Department of Neuropathology, Emory University Hospital, for the histology and immunohistochemistry analyses. The laboratory of D.M. Desiderio contributed the proteomic data, and the laboratory of N.M. Oyesiku contributed the tumor samples, microarray and reverse transcriptase- real time quantitative PCR data to this study.

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Authors and Affiliations

  1. Department of Neurosurgery and Laboratory of Molecular Neurosurgery and Biotechnology, Emory University School of Medicine, 1365 B Clifton Rd., NE, Suite. 6200, Atlanta, GA, 30322, USA

    Chheng-Orn Evans & Nelson M. Oyesiku

  2. Department of Pathology and Laboratory Medicine and Winship Cancer Institute, Emory University School of Medicine, Atlanta, GA, USA

    Carlos S. Moreno

  3. Charles B. Stout Neuroscience Mass Spectrometry Laboratory, University of Tennessee, Health Science Center, Knoxville, TN, USA

    Xianquan Zhan & Dominic M. Desiderio

  4. Department of Radiation Oncology, Winship Cancer Institute, Emory University School of Medicine, Atlanta, GA, USA

    Michael T. McCabe & Paula M. Vertino

  5. Department of Neurology, University of Tennessee, Health Science Center, Knoxville, TN, USA

    Dominic M. Desiderio

  6. Department of Molecular Sciences, University of Tennessee, Health Science Center, Knoxville, TN, USA

    Dominic M. Desiderio

  7. Cancer Institute, University of Tennessee, Health Science Center, Knoxville, TN, USA

    Dominic M. Desiderio

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  1. Chheng-Orn Evans
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  2. Carlos S. Moreno
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  3. Xianquan Zhan
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  4. Michael T. McCabe
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  5. Paula M. Vertino
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  6. Dominic M. Desiderio
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  7. Nelson M. Oyesiku
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Correspondence to Nelson M. Oyesiku.

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Fig. S2

Methylation Specific PCR Analysis of Normal Pituitary and Prolactinoma Samples. DNA isolated from eight normal pituitaries and nine prolactinomas was analyzed by methylation specific PCR for HLAG and GSTP1 CpG islands. Controls included a sample from a primary normal lung fibroblast (Unmeth) and genomic DNA that was in vitro methylated with the M.SssI DNA methyltransferase (Meth). U, unmethylated primer set. M, methylated primer set (ppt 109 kb)

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Evans, CO., Moreno, C.S., Zhan, X. et al. Molecular pathogenesis of human prolactinomas identified by gene expression profiling, RT-qPCR, and proteomic analyses. Pituitary 11, 231–245 (2008). https://doi.org/10.1007/s11102-007-0082-2

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