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Quantitative real-time PCR does not show selective targeting of p14ARF but concomitant inactivation of both p16INK4A and p14ARF in 105 human primary gliomas

Abstract

In many human cancers, the INK4A locus is frequently mutated by homozygous deletions. By alternative splicing this locus encodes two non-related tumor suppressor genes, p16INK4A and p14ARF (p19ARF in mice), which regulate cell cycle and cell survival in the retinoblastoma protein (pRb) and p53 pathways, respectively. In mice, the role of p16INK4A as the critical tumor suppressor gene at the INK4A locus was challenged when it was found that p19ARF only knock-out mice developed tumors, including gliomas. We have analysed the genetic status of the INK4A locus in 105 primary gliomas using both microsatellite mapping (MSM) and quantitative real-time PCR (QRT–PCR). Comparison of the results of the two methods revealed agreement in 67% of the tumors examined. In discordant cases, fluorescence in situ hybridization (FISH) analysis was always found to support QRT–PCR classification. Direct assessment of p14ARF exon 1β, p16INK4A exon 1α and exon 2 by QRT–PCR revealed 43 (41%) homozygous and eight (7%) hemizygous deletions at the INK4A locus. In 49 (47%) gliomas, both alleles were retained. In addition, QRT–PCR, but not MSM, detected hyperploidy in five (5%) tumors. Deletion of p14ARF was always associated with co-deletion of p16INK4A and increased in frequency upon progression from low to high grade gliomas. Shorter survival was associated with homozygous deletions of INK4A in the subgroup of glioblastoma patients older than 50 years of age (P=0.025, Anova test single factor, α=0.05).

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Acknowledgements

We would like to thank Annie-Claire Diserens and Marie-France Hamou for technical assistance. Supported by grants from the Swiss National Science Foundation (No. 31053746.98, to A Merlo; No. 3153625.98 to EJM Speel; No. 3149194.96 to EG Van Meir) and the Swiss Cancer League (No. 614-2-1998 to A Merlo; No. 649-2-1998 to EJM Speel; No. 728-9-1998 and No. 904-09-1999 to ME Hegi).

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Labuhn, M., Jones, G., Speel, E. et al. Quantitative real-time PCR does not show selective targeting of p14ARF but concomitant inactivation of both p16INK4A and p14ARF in 105 human primary gliomas. Oncogene 20, 1103–1109 (2001). https://doi.org/10.1038/sj.onc.1204197

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