Development of cross-resistance between heat and cisplatin or hydroxyurea treatments in FaDu squamous carcinoma cells.
BACKGROUND
Induction of hyperthermia by radiofrequency ablation is gaining popularity in treating a variety of solid tumors. This study examined an impact of sublethal heat treatment interacted with chemotherapeutic drugs on the survival of head and neck squamous carcinoma cells using in vitro model.


METHODS
FaDu cells were subjected to heat treatment at 42 degrees C or 45 degrees C for 15 min either before or after exposure to cisplatin or hydroxyurea. The survival of cells was determined by 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide assay. The RNA and protein levels of various heat shock proteins were examined by reverse transcription polymerase chain reaction and Western blot analysis, respectively. Cell cycle progression was analyzed by flow cytometry with propidium iodide staining.


RESULTS
FaDu cells preheated to 45 degrees C exhibited an increased resistance to hydroxyurea but not to cisplatin. The heat treatment resulted in induction of HSP70 expression at transcript and protein levels, but there was no change in expression of HSP90beta and HSP27. After heat treatment, cells accumulated in S-phase at 3 h and proceeded to G(2)/M phase at 24 h. When cells pre-exposed to drugs for 24 h, the cisplatin-treated cells exhibited a higher thermotolerance than the hydroxyurea-treated cells at heat treatment of 45 degrees C. Cisplatin and hydroxyurea caused cells to accumulate in S-phase and increased the protein expression of HSP27 but not HSP90beta and HSP70.


CONCLUSIONS
FaDu cells surviving the heat treatment expressed HSP70 and disrupted cell cycle progression, which resulted in developing a resistance to subsequent hydroxyurea treatment. However, the heat treatment did not have an effect on the sensitivity to cisplatin. In the reversed procedure, pre-exposure to hydroxyurea and cisplatin resulted in developing a thermotolerance.
Yang
Weng-Lang
WL
Department of Surgery, Albert Einstein College of Medicine and Montefiore Medical Center, Bronx, New York 10461, USA. wlyang@aecom.yu.edu
Yang-Biggs
Grace
G
Wu
Yin
Y
Ye
Xuemei
X
Gallos
George
G
Owen
Randall P
RP
Ravikumar
T S
TS
D000970:Antineoplastic Agents / Q000008:administration & dosage; D002294:Carcinoma, Squamous Cell / Q000737:chemistry / Q000473:pathology / Q000628:therapy*; D002470:Cell Survival; D002945:Cisplatin / Q000008:administration & dosage*; D019008:Drug Resistance, Neoplasm; D016195:G2 Phase; D055551:HSP27 Heat-Shock Proteins; D018840:HSP70 Heat-Shock Proteins / Q000032:analysis / Q000235:genetics; D018841:HSP90 Heat-Shock Proteins / Q000032:analysis / Q000235:genetics; D006258:Head and Neck Neoplasms / Q000737:chemistry / Q000473:pathology / Q000628:therapy*; D006360:Heat-Shock Proteins; D006358:Hot Temperature; D006801:Humans; D006918:Hydroxyurea / Q000008:administration & dosage*; D007700:Kinetics; D008938:Mitosis; D018832:Molecular Chaperones; D009363:Neoplasm Proteins / Q000032:analysis / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016196:S Phase; D014407:Tumor Cells, Cultured
D000970:Antineoplastic Agents; D055551:HSP27 Heat-Shock Proteins; D018840:HSP70 Heat-Shock Proteins; D018841:HSP90 Heat-Shock Proteins; C057725:HSPB1 protein, human; D006360:Heat-Shock Proteins; D018832:Molecular Chaperones; D009363:Neoplasm Proteins; D012333:RNA, Messenger; D002945:Cisplatin; D006918:Hydroxyurea