PMID- 16945015
OWN - NLM
STAT- MEDLINE
DCOM- 20070112
LR  - 20131121
IS  - 1540-658X (Print)
IS  - 1540-658X (Linking)
VI  - 4
IP  - 4
DP  - 2006 Aug
TI  - Quantitative characterization of mitosis-blocked tetraploid cells using high 
      content analysis.
PG  - 421-42
AB  - A range of cellular evidence supporting a G1 tetraploidy checkpoint was obtained 
      from different assay methods including flow cytometry, immunoblotting, and 
      microscopy. Cancer research would benefit if these cellular properties could 
      instead be measured by a single, quantitative, automated assay method, such as 
      high content analysis (HCA). Thus, nocodazole-treated cells were fluorescently 
      labeled for different cell cycle-associated properties, including DNA content, 
      retinoblastoma (Rb) and histone H3 phosphorylation, p53 and p21(WAF1) expression, 
      nuclear and cell sizes, and cell morphology, and automatically imaged, analyzed, 
      and correlated using HCA. HCA verified that nocodazole-induced mitosis block 
      resulted in tetraploid cells. Rb and histone H3 were maximally 
      hyperphosphorylated by 24 h of nocodazole treatment, accompanied by cell and 
      nuclear size decreases and cellular rounding. Cells remained tetraploid and 
      mononucleated with longer treatments, but other targets reverted to G1 levels, 
      including Rb and histone H3 dephosphorylation accompanied by cellular 
      respreading. This was accompanied by increased p53 and p21(WAF1) expression 
      levels. The range of effects accompanying nocodazole-induced block of mitosis and 
      the resulting tetraploid cells' reversal to a pseudo-G1 state can be 
      quantitatively measured by HCA in an automated manner, recommending this assay 
      method for the large-scale biology challenges of modern cancer drug discovery.
FAU - Grove, Linnette E
AU  - Grove LE
AD  - Cellomics, Inc., Pittsburgh, PA, USA.
FAU - Ghosh, Richik N
AU  - Ghosh RN
LA  - eng
PT  - Journal Article
PL  - United States
TA  - Assay Drug Dev Technol
JT  - Assay and drug development technologies
JID - 101151468
RN  - 0 (Antineoplastic Agents)
RN  - 0 (Cell Cycle Proteins)
RN  - 0 (Cyclin-Dependent Kinase Inhibitor p21)
RN  - 0 (DNA, Neoplasm)
RN  - 0 (Histones)
RN  - 0 (Retinoblastoma Protein)
RN  - 0 (Tumor Suppressor Protein p53)
RN  - SH1WY3R615 (Nocodazole)
SB  - IM
MH  - Antineoplastic Agents/chemical synthesis
MH  - Cell Cycle Proteins/*analysis/metabolism
MH  - Cell Line, Tumor
MH  - Cell Nucleus/drug effects/metabolism
MH  - Cell Proliferation/drug effects
MH  - Cell Shape/drug effects
MH  - Cell Size/drug effects
MH  - Cyclin-Dependent Kinase Inhibitor p21/analysis/metabolism
MH  - DNA, Neoplasm/*analysis/genetics/metabolism
MH  - Dose-Response Relationship, Drug
MH  - Drug Design
MH  - Histones/analysis/metabolism
MH  - Humans
MH  - Image Interpretation, Computer-Assisted/instrumentation/methods
MH  - Mitosis/*drug effects/genetics
MH  - Mitotic Index
MH  - Nocodazole/pharmacology
MH  - Phosphorylation
MH  - *Polyploidy
MH  - Retinoblastoma Protein/analysis/metabolism
MH  - Technology, Pharmaceutical/instrumentation/methods
MH  - Time Factors
MH  - Tumor Suppressor Protein p53/analysis/metabolism
EDAT- 2006/09/02 09:00
MHDA- 2007/01/16 09:00
CRDT- 2006/09/02 09:00
PHST- 2006/09/02 09:00 [pubmed]
PHST- 2007/01/16 09:00 [medline]
PHST- 2006/09/02 09:00 [entrez]
AID - 10.1089/adt.2006.4.421 [doi]
PST - ppublish
SO  - Assay Drug Dev Technol. 2006 Aug;4(4):421-42. doi: 10.1089/adt.2006.4.421.