PMID- 9291941
OWN - NLM
STAT- MEDLINE
DCOM- 19971015
LR  - 20190512
IS  - 0022-3069 (Print)
IS  - 0022-3069 (Linking)
VI  - 56
IP  - 9
DP  - 1997 Sep
TI  - Detection of p16 gene deletions in gliomas: a comparison of fluorescence in situ 
      hybridization (FISH) versus quantitative PCR.
PG  - 999-1008
AB  - The p16 protein plays a key role in cell cycle control by preventing CDK4 from 
      inactivating the retinoblastoma protein (pRb). The corresponding tumor suppressor 
      gene (p16/MTS1/CDKN2) has recently been implicated in malignant progression of 
      astrocytomas and could potentially serve as an important marker for patient 
      prognosis and for guiding specific therapeutic strategies. We have undertaken a 
      study to evaluate 2 methods of detecting p16 deletion. Thirty diffuse gliomas 
      were analyzed for p16 gene dosage. Dual color fluorescence in situ hybridization 
      (FISH) was performed on cytologic preparations using paired centromeric (CEN) and 
      locus-specific probes for CEN9/p16, CEN8/RB, and CEN12/CDK4. Quantitative PCR was 
      performed using primers for p16, MTAP, and reference genes. Eleven cases were 
      also studied using comparative genomic hybridization (CGH). Abnormalities of the 
      p16-CDK4-RB pathway were identified in 21 (70%) cases by FISH and/or PCR. These 
      included 15 (50%) with p16 deletion, 9 of which were detected by both techniques, 
      3 by FISH alone, and 3 by PCR alone (concordance rate = 81%). FISH analysis 
      further revealed tetraploidy/aneuploidy in 14 (47%), RB deletion in 11 (37%) and 
      CDK4 amplification in 1 (3.3%). There were 94% and 100% concordance rates between 
      CGH and FISH or PCR, respectively. Quantitative PCR was noninformative in 4 
      cases. Although FISH and quantitative PCR are both reliable techniques, each has 
      limitations. PCR is likely to miss p16 deletions when there is significant normal 
      cell contamination or clonal heterogeneity, whereas the p16 YAC probe used for 
      FISH analysis may miss small deletions. Replacement of the latter with a cosmid 
      probe may improve the sensitivity of FISH in future experiments.
FAU - Perry, A
AU  - Perry A
AD  - Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, 
      Minnesota 55905, USA.
FAU - Nobori, T
AU  - Nobori T
FAU - Ru, N
AU  - Ru N
FAU - Anderl, K
AU  - Anderl K
FAU - Borell, T J
AU  - Borell TJ
FAU - Mohapatra, G
AU  - Mohapatra G
FAU - Feuerstein, B G
AU  - Feuerstein BG
FAU - Jenkins, R B
AU  - Jenkins RB
FAU - Carson, D A
AU  - Carson DA
LA  - eng
GR  - CA 64976/CA/NCI NIH HHS/United States
GR  - CA50905/CA/NCI NIH HHS/United States
GR  - CA64898/CA/NCI NIH HHS/United States
PT  - Comparative Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - England
TA  - J Neuropathol Exp Neurol
JT  - Journal of neuropathology and experimental neurology
JID - 2985192R
RN  - 0 (Carrier Proteins)
RN  - 0 (Cyclin-Dependent Kinase Inhibitor p16)
SB  - IM
MH  - Brain Neoplasms/*genetics
MH  - Carrier Proteins/*genetics
MH  - Cyclin-Dependent Kinase Inhibitor p16
MH  - *Gene Deletion
MH  - Glioma/*genetics
MH  - Humans
MH  - In Situ Hybridization, Fluorescence
MH  - Nucleic Acid Hybridization
MH  - Polymerase Chain Reaction
EDAT- 1997/09/18 00:00
MHDA- 1997/09/18 00:01
CRDT- 1997/09/18 00:00
PHST- 1997/09/18 00:00 [pubmed]
PHST- 1997/09/18 00:01 [medline]
PHST- 1997/09/18 00:00 [entrez]
AID - 10.1097/00005072-199709000-00005 [doi]
PST - ppublish
SO  - J Neuropathol Exp Neurol. 1997 Sep;56(9):999-1008. doi: 
      10.1097/00005072-199709000-00005.