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Abrogation of the Rb/p16 Tumor-suppressive Pathway in Virtually All Pancreatic Carcinomas¹

Departments of Oncology [M. S., R. H. H., S. B. B., S. E. K., J. G. H.] and Pathology [R. H. H., W. H., C. A. M., S. E. K.], The Johns Hopkins Medical Institutions, Baltimore, Maryland 21205; Department of Pathology and Laboratory Medicine, University of North Carolina School of Medicine, Chapel Hill, North Carolina 27599 [J. G., R. M.]; Oncology/Immunology Research, Wyeth Ayerst Research, Pearl River, New York 10965 [S. K. R.]; and Medical Clinic IMBL, Ruhr-University Bochum, Bochum, Germany [S. A. H., I. S. W., W. S.]

The Rb/p16 tumor-suppressive pathway is abrogated frequently in human tumors, either through inactivation of the Rb or p16^{INK4a/CDKN2/MTS1} tumor-suppressor proteins, or through alteration or overexpression of the cyclin D1 or cyclin-dependent kinase 4 oncoproteins. We reported previously that the p16 gene was genetically inactivated in 82% of pancreatic carcinomas. Nearly half of these inactivations were by intragenic mutation of p16, and the remainder were by homozygous deletion of the gene. Here, we analyzed pancreatic carcinomas for additional mechanisms by which the Rb/p16 pathway might be inactivated.

Transcriptional silencing of the p16 gene in association with methylation of its 5'-CpG island was examined by methylation-specific PCR in 18 pancreatic carcinomas. Nine of these were known to harbor an intragenic mutation in p16, and nine had a wild-type p16 coding sequence. Seven of the 18 tumors were hypermethylated, and all 7 were p16 wild-type (P = 0.001). Complete silencing of transcription from methylated wild-type gene sequences was demonstrated. Immunohistochemical analysis revealed normal expression levels of the Rb protein in all carcinomas studied. None of the carcinomas had genomic amplification of the cyclin D1 or CDK4 genes, and none had mutation of the p16-binding domain of CDK4. An additional p16 mutation was identified.

In total, the Rb/p16 pathway was abrogated in 49 of the 50 carcinomas (98%) studied, all through inactivation of the p16 gene. Similar results were obtained in an independently analyzed series of 19 pancreatic carcinomas. These data demonstrate the central role of the Rb/p16 pathway in the development of pancreatic carcinoma.

¹ This work was supported by the Specialized Programs of Research Excellence in Gastrointestinal Cancer and Lung Cancer (NIH Grants CA62924 and CA58184, respectively), by the Deutsche Krebshilfe (Grant 10-1137-Hal), and by the Ruhr-Universität Bochum (Grant RUB-Med-FO-808004). S. E. K. is a McDonnell Foundation Scholar. S. B. B. and J. G. H. receive research funding and are entitled to sales royalties from ONCOR, which is developing products related to research described in this paper. The terms of this arrangement have been reviewed and approved by The Johns Hopkins University in accordance with its conflict of interest policies.

² To whom requests for reprints should be addressed, at Department of Oncology, The Johns Hopkins Medical Institutions, Baltimore, MD 21205-2196. Phone: (410) 614-3314; Fax: (410) 614-0671.

Received 5/ 6/97. Accepted 6/12/97.

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				S. Kobayashi, H. Shirasawa, H. Sashiyama, H. Kawahira, K. Kaneko, T. Asano, and T. Ochiai p16INK4a Expression Adenovirus Vector to Suppress Pancreas Cancer Cell Proliferation Clin. Cancer Res., December 1, 1999; 5(12): 4182 - 4185. [Abstract] [Full Text] [PDF]

				L. H. Ellenson hMLH1 Promoter Hypermethylation in Microsatellite Instability-Positive Endometrial Carcinoma : Cause or Consequence? Am. J. Pathol., November 1, 1999; 155(5): 1399 - 1402. [Full Text] [PDF]

				D. J. Wong, S. A. Foster, D. A. Galloway, and B. J. Reid Progressive Region-Specific De Novo Methylation of the p16 CpG Island in Primary Human Mammary Epithelial Cell Strains during Escape from M0 Growth Arrest Mol. Cell. Biol., August 1, 1999; 19(8): 5642 - 5651. [Abstract] [Full Text] [PDF]

				R. Z. Stolzenberg-Solomon, D. Albanes, F. J. Nieto, T. J. Hartman, J. A. Tangrea, M. Rautalahti, J. Sehlub, J. Virtamo, and P. R. Taylor Pancreatic Cancer Risk and Nutrition-Related Methyl-Group Availability Indicators in Male Smokers J Natl Cancer Inst, March 17, 1999; 91(6): 535 - 541. [Abstract] [Full Text] [PDF]

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				N. Liu, T. Furukawa, M. Kobari, and M.-S. Tsao Comparative Phenotypic Studies of Duct Epithelial Cell Lines Derived from Normal Human Pancreas and Pancreatic Carcinoma Am. J. Pathol., July 1, 1998; 153(1): 263 - 269. [Abstract] [Full Text] [PDF]