PMID- 9609099
OWN - NLM
STAT- MEDLINE
DCOM- 19980616
LR  - 20191024
IS  - 0899-1987 (Print)
IS  - 0899-1987 (Linking)
VI  - 22
IP  - 1
DP  - 1998 May
TI  - Mapping a novel cellular-senescence gene to human chromosome 2q37 by irradiation 
      microcell-mediated chromosome transfer.
PG  - 34-45
AB  - To identify the subchromosomal region that carries the 
      cellular-senescence-restoring program of the human cervical carcinoma cell line 
      SiHa, we constructed by irradiation microcell-mediated chromosome transfer a 
      library of mouse A9 cells containing various fragments of human chromosome 2 
      tagged with pSV2neo in 2p11-p12. Eighty-seven clones were isolated and screened 
      for the presence of human sequences by inter-Alu and inter-L1 polymerase chain 
      reaction (PCR), and six clones exhibiting PCR-laddering patterns that differed 
      from those of the A9 cells containing an intact chromosome 2 were examined 
      further. Chromosome analysis and fluorescence in situ hybridization (FISH) using 
      human-specific repetitive sequences revealed that four of these clones contained 
      single subchromosomal transferable fragments (STFs). Southern blot hybridization 
      of 14 cosmid markers revealed that the STFs in A9 cells were derived from human 
      chromosome 2. These STFs were transferred into SiHa cells by microcell fusion, 
      and one of the STFs restored the cellular-senescence program. The concordance of 
      the cellular-senescence-restoring program with the presence or absence of 
      specific DNA fragments of chromosome 2 indicated that the putative 
      cellular-senescence gene was located in 2q32-qter. For more detailed mapping, we 
      constructed mouse A9 cells containing STFs derived from human chromosome 2 tagged 
      with pSTneo at different regions in 2q31-qter. PCR-laddering and FISH analyses 
      were used to identify six clones that contained different STFs. These STFs were 
      transferred into SiHa cells, and one of the three clones that restored cellular 
      senescence contained a small fragment of human chromosome 2. This STF was shown 
      by PCR analysis using 14 human chromosome 2-specific primer pairs to be smaller 
      than 12.2 cM and was mapped to the 2q37 region by FISH analysis with inter-Alu 
      PCR. Beta-galactosidase activity, which is a biomarker of senescent cells, and 
      telomerase activity similar to that found in parental SiHa cells were detected in 
      SiHa microcell hybrids, suggesting that the putative cellular-senescence gene was 
      not involved in a telomerase pathway but rather in an alternate pathway of 
      cellular senescence.
FAU - Uejima, H
AU  - Uejima H
AD  - Department of Molecular and Cell Genetics, School of Life Sciences, Faculty of 
      Medicine, Tottori University, and CREST (JST), Yonago, Japan.
FAU - Shinohara, T
AU  - Shinohara T
FAU - Nakayama, Y
AU  - Nakayama Y
FAU - Kugoh, H
AU  - Kugoh H
FAU - Oshimura, M
AU  - Oshimura M
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Mol Carcinog
JT  - Molecular carcinogenesis
JID - 8811105
RN  - 0 (DNA Primers)
RN  - 0 (Fluorescent Dyes)
RN  - 0 (Genetic Markers)
SB  - IM
MH  - Animals
MH  - Cell Line
MH  - Cellular Senescence/*genetics
MH  - Chromosome Mapping/*methods
MH  - *Chromosomes, Human, Pair 2
MH  - Cosmids
MH  - DNA Primers
MH  - Female
MH  - Fluorescent Dyes
MH  - *Gene Transfer Techniques
MH  - Genetic Markers
MH  - Humans
MH  - In Situ Hybridization, Fluorescence/methods
MH  - Karyotyping
MH  - Mice
MH  - Polymerase Chain Reaction/methods
MH  - Sequence Tagged Sites
MH  - Tumor Cells, Cultured
MH  - Uterine Cervical Neoplasms
EDAT- 1998/06/03 00:00
MHDA- 1998/06/03 00:01
CRDT- 1998/06/03 00:00
PHST- 1998/06/03 00:00 [pubmed]
PHST- 1998/06/03 00:01 [medline]
PHST- 1998/06/03 00:00 [entrez]
AID - 10.1002/(SICI)1098-2744(199805)22:1<34::AID-MC5>3.0.CO;2-L [pii]
AID - 10.1002/(sici)1098-2744(199805)22:1<34::aid-mc5>3.0.co;2-l [doi]
PST - ppublish
SO  - Mol Carcinog. 1998 May;22(1):34-45. doi: 
      10.1002/(sici)1098-2744(199805)22:1<34::aid-mc5>3.0.co;2-l.