PMID- 12438006
OWN - NLM
STAT- MEDLINE
DCOM- 20030117
LR  - 20190702
IS  - 0027-5107 (Print)
IS  - 0027-5107 (Linking)
VI  - 521
IP  - 1-2
DP  - 2002 Nov 26
TI  - Enhancing the in vitro and in vivo detection of aneuploidy by fluorescence in 
      situ hybridization with the use of bromodeoxyuridine as a proliferation marker.
PG  - 81-9
AB  - Aneuploidy is associated with spontaneous abortions, birth defects, and many 
      types of human cancers. Currently there are few assays developed for the 
      efficient detection of aneuploidy in vivo. However, with the recent availability 
      of chromosome-specific DNA probes for the rat, fluorescence in situ hybridization 
      (FISH) techniques could be used for the rapid and sensitive detection of 
      aneuploidy in different tissue and cell types. In order to develop a system that 
      can detect alterations in chromosome number in rat cells in vitro, we treated 
      cultured rat lymphocytes with three aneugens-noscapine hydrochloride (0-150 
      microM) and vincristine and vinblastine sulfate (0-0.06 microM). 
      5-Bromo-2-deoxyuridine (BrdU; 1 microM) was added to the culture medium to allow 
      proliferating and non-proliferating cells to be distinguished. To test this assay 
      under in vivo conditions, 21-day-old male Sprague-Dawley rats were subcutaneously 
      implanted with osmotic pumps that delivered BrdU (approximately 12 mg/kg per day) 
      continuously. These rats were administered vinblastine sulfate (0, 0.5 and 
      1mg/kg) by intraperitoneal injection. The rat lymphocytes and hepatocytes 
      incorporating BrdU were detected by immuno-fluorescent labeling, and FISH with a 
      rat chromosome 4 probe was performed on the labeled and unlabeled cells. Highly 
      significant increases in hyperdiploidy were seen in the replicating rat 
      lymphocytes treated with noscapine, vincristine or vinblastine in vitro and in 
      the rat hepatocytes treated with vinblastine in vivo. In contrast, no significant 
      increase in hyperdiploidy was observed in the non-replicating cells. These 
      results demonstrate that this BrdU-enhanced FISH assay with chromosome-specific 
      rat probes can be used to efficiently detect numerical chromosomal aberrations in 
      vitro and in vivo in slowly or moderately replicating rat tissues. The 
      combination of BrdU-labeling and FISH allows the scoring of hyperdiploidy to be 
      focused on the actively replicating cells, thereby increasing the sensitivity of 
      the FISH technique.
FAU - Balakrishnan, S
AU  - Balakrishnan S
AD  - Environmental Toxicology Graduate Program, University of California, Riverside, 
      CA 92521, USA.
FAU - Payawal, J
AU  - Payawal J
FAU - Schuler, M J
AU  - Schuler MJ
FAU - Hasegawa, L
AU  - Hasegawa L
FAU - Eastmond, D A
AU  - Eastmond DA
LA  - eng
PT  - Journal Article
PT  - Research Support, U.S. Gov't, Non-P.H.S.
PL  - Netherlands
TA  - Mutat Res
JT  - Mutation research
JID - 0400763
RN  - 0 (Biomarkers)
RN  - 5J49Q6B70F (Vincristine)
RN  - 5V9KLZ54CY (Vinblastine)
RN  - 8V32U4AOQU (Noscapine)
RN  - G34N38R2N1 (Bromodeoxyuridine)
SB  - IM
MH  - *Aneuploidy
MH  - Animals
MH  - Biomarkers
MH  - Bromodeoxyuridine/analysis/*metabolism
MH  - Cell Division/drug effects
MH  - Cells, Cultured
MH  - Hepatocytes/drug effects/physiology
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Lymphocytes/drug effects/physiology
MH  - Male
MH  - Noscapine/adverse effects
MH  - Rats
MH  - Rats, Inbred F344
MH  - Rats, Sprague-Dawley
MH  - Sensitivity and Specificity
MH  - Vinblastine/adverse effects
MH  - Vincristine/adverse effects
EDAT- 2002/11/20 04:00
MHDA- 2003/01/18 04:00
CRDT- 2002/11/20 04:00
PHST- 2002/11/20 04:00 [pubmed]
PHST- 2003/01/18 04:00 [medline]
PHST- 2002/11/20 04:00 [entrez]
AID - S1383571802002206 [pii]
AID - 10.1016/s1383-5718(02)00220-6 [doi]
PST - ppublish
SO  - Mutat Res. 2002 Nov 26;521(1-2):81-9. doi: 10.1016/s1383-5718(02)00220-6.