PMID- 16228978
OWN - NLM
STAT- MEDLINE
DCOM- 20060203
LR  - 20121115
IS  - 1552-4922 (Print)
IS  - 1552-4922 (Linking)
VI  - 68
IP  - 2
DP  - 2005 Dec
TI  - Analysis of hypoxia-inducible factor-1alpha accumulation and cell cycle in 
      geldanamycin-treated human cervical carcinoma cells by laser scanning cytometry.
PG  - 59-70
AB  - BACKGROUND: Tumor hypoxia has been linked to increased disease aggressiveness and 
      poorer treatment outcomes, and the transcription factor hypoxia-inducible 
      factor-1 (HIF-1) has been identified as the key molecule mediating the cellular 
      response to hypoxic microenvironments. The alpha-subunit of this factor is 
      accumulated under hypoxia and rapidly degraded during re-oxygenation, rendering 
      the reliable measurement of HIF-1alpha a difficult task. Heat shock protein 90 
      (Hsp90) is an essential protein that controls the activity, turnover, and 
      trafficking of a variety of other proteins including HIF-1alpha and cell cycle 
      regulators. Hsp90 inhibitors like geldanamycin therefore have the potential to 
      target tumor-cell survival by at least two mechanisms, compromising the 
      accumulation of HIF-1alpha and cell proliferation. METHODS: We describe here the 
      simultaneous measurement of HIF-1alpha and cell cycle parameters by laser 
      scanning cytometry (LSC) after exposure of two different human cervical carcinoma 
      cell lines to hypoxia and geldanamycin. RESULTS: Our analysis demonstrates that 
      the cell lines react to hypoxia and drug treatment in a distinct way, with SiHa 
      being more affected by low oxygen concentrations than is ME180, which was more 
      sensitive to geldanamycin treatment. Both cell lines respond to geldanamycin with 
      a G(2)/M-phase arrest and a decrease in HIF-1alpha accumulation. Cell death due 
      to geldanamycin occurs in association with mitosis, presumably through mitotic 
      catastrophe. CONCLUSION: Our results indicate that LSC can significantly 
      contribute to the evaluation of in vitro drug effects particularly with respect 
      to tumor hypoxia and the measurement of HIF-1alpha.
FAU - Schwock, Jorg
AU  - Schwock J
AD  - Department of Laboratory Medicine and Pathobiology, University of Toronto, 
      Toronto, Ontario, Canada.
FAU - Geddie, William R
AU  - Geddie WR
FAU - Hedley, David W
AU  - Hedley DW
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Cytometry A
JT  - Cytometry. Part A : the journal of the International Society for Analytical 
      Cytology
JID - 101235694
RN  - 0 (Antibiotics, Antineoplastic)
RN  - 0 (Benzoquinones)
RN  - 0 (HIF1A protein, human)
RN  - 0 (HSP90 Heat-Shock Proteins)
RN  - 0 (Hypoxia-Inducible Factor 1, alpha Subunit)
RN  - 0 (Lactams, Macrocyclic)
RN  - 0 (Quinones)
RN  - Z3K3VJ16KU (geldanamycin)
SB  - IM
MH  - Antibiotics, Antineoplastic/*pharmacology
MH  - Benzoquinones
MH  - Blotting, Western
MH  - Cell Cycle/drug effects
MH  - Cell Hypoxia
MH  - Cell Line, Tumor
MH  - Cell Proliferation/drug effects
MH  - Female
MH  - Fluorescent Antibody Technique
MH  - G2 Phase/drug effects
MH  - HSP90 Heat-Shock Proteins/antagonists & inhibitors/physiology
MH  - Humans
MH  - Hypoxia-Inducible Factor 1, alpha Subunit/*analysis
MH  - Lactams, Macrocyclic
MH  - *Laser Scanning Cytometry
MH  - Mitosis/drug effects
MH  - Quinones/*pharmacology
MH  - Uterine Cervical Neoplasms/*chemistry/*pathology/physiopathology
EDAT- 2005/10/18 09:00
MHDA- 2006/02/04 09:00
CRDT- 2005/10/18 09:00
PHST- 2005/10/18 09:00 [pubmed]
PHST- 2006/02/04 09:00 [medline]
PHST- 2005/10/18 09:00 [entrez]
AID - 10.1002/cyto.a.20192 [doi]
PST - ppublish
SO  - Cytometry A. 2005 Dec;68(2):59-70. doi: 10.1002/cyto.a.20192.