Document Detail

Expression of proliferation associated antigens in the cell cycle of synchronized mammalian cells.
MedLine Citation:
PMID:  1372207     Owner:  NLM     Status:  MEDLINE    
Flow cytometric bivariate analysis was used to investigate the expression of PCNA, p120 and p145 during the cell cycle of a mammalian cell line (CHO-K1). Initially, aliquots of cells in exponential and plateau (G0) phase were analyzed for proliferation associated antigen expression. Expression of PCNA and p145 during G0 was markedly depressed (less than 12% positive) while 54% of the G0 cells stained positive for p120. The fluorescent intensity (mean channel fluorescence) of these G0 positive p120 cells, however, was only slightly above the mean channel fluorescence (MCF) of cells stained with a negative isotype control. In asynchronous cultures, all three antigens were expressed in greater than 70% of the cells, with PCNA staining being greater than 95%. Cells were then synchronized using mitotic selection (mitotic index of 97%) and antigen levels were measured as cells progressed synchronously through the cell cycle. From DNA analysis histograms, it appeared that the degree of synchrony was approximately 90% throughout the remainder of the cell cycle. The bivariate DNA/PCNA, DNA/p120, and DNA/p145 histograms for mitotic cells indicated that both p120 and p145 expression were elevated (percent positive and MCF) while PCNA levels were near controls (MCF). In early G1, all three markers were depressed (less than 12% positive); however PCNA levels rose precipitously in mid-G1 (greater than 50% positive). In late G1 to early S, p145 levels increased concomitantly with increases in p120. All three antigens were elevated throughout S phase and began to decline as cells moved from G2/M to G1 of the next cell cycle with p145 expression decreasing first. This report indicates that all three proliferation associated antigens studied are differentially expressed in the cell cycle and therefore may be useful in detecting and assessing the proliferation state.
W E Bolton; W R Mikulka; C G Healy; R J Schmittling; N S Kenyon
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Cytometry     Volume:  13     ISSN:  0196-4763     ISO Abbreviation:  Cytometry     Publication Date:  1992  
Date Detail:
Created Date:  1992-04-17     Completed Date:  1992-04-17     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  8102328     Medline TA:  Cytometry     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  117-26     Citation Subset:  IM    
Coulter Immunology, Division of Coulter Corporation, Hialeah, Florida 33010.
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MeSH Terms
CHO Cells / immunology
Cell Division / physiology
Cells, Cultured
DNA / analysis,  genetics
Flow Cytometry
G0 Phase / physiology
G1 Phase / physiology
Gene Expression / physiology
Nuclear Proteins / analysis*,  genetics
Proliferating Cell Nuclear Antigen
S Phase
Staining and Labeling
Reg. No./Substance:
0/Nuclear Proteins; 0/Proliferating Cell Nuclear Antigen; 9007-49-2/DNA

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