Effects of 17-AAG on the cell cycle and apoptosis of H446 cells and the associated mechanisms.
As a heat shock protein 90 inhibitor, 17-allylamino-17‑demethoxygeldanamycin (17-AAG) has been studied in numerous types of cancer, however the effects of 17-AAG on apoptosis and the cell cycle of H446 cells remain unclear. In the current study, the MTT method was used to evaluate the inhibitory effects of different durations and doses of 17‑AAG treatment on the proliferation of H446 cells. The cells were stained with Annexin-fluorescein isothiocyanate/propidium iodide and measured by flow cytometry, and the gene and protein expression levels of signal transducer and activator of transcription 3 (STAT3), survivin, cyclin D1, cyt‑C, caspase 9 and caspase 3 were determined by reverse transcription‑quantitative polymerase chain reaction and western blot analysis. The results indicated that with treatment with 1.25‑20 mg/l 17‑AAG for 24 and 48 h, significant inhibition of H446 cell proliferation was observed in a time‑ and dose‑dependent manner. With treatment of 3.125, 6.25 and 12.5 mg/l 17‑AAG for 48 h, significant apoptosis and cell cycle arrest was observed. The results indicated that the gene and protein expression levels of STAT3, survivin and cyclin D1 were downregulated, and cyt‑C, caspase 9 and caspase 3 were upregulated by 17‑AAG in a dose-dependent manner when the cells were treated with 3.125 and 6.25 mg/l 17-AAG for 48 h. The results indicated that 17‑AAG is able to inhibit the cell proliferation, induce apoptosis and G2/M arrest and downregulate the gene and protein expression levels of STAT3, survivin and cyclin D1, and upregulate gene and protein expression of cyt‑C, caspase 9, caspase 3.
Department of Immunology, Chengde Medical University, Chengde, Hebei 067000, P.R. China.
Xuerong
X
Zhao
Department of Immunology, Chengde Medical University, Chengde, Hebei 067000, P.R. China.
Jianping
J
Wang
Department of Immunology, Chengde Medical University, Chengde, Hebei 067000, P.R. China.
Lijun
L
Xiao
Department of Fundamental Research, Chengde Medical University, Chengde, Hebei 067000, P.R. China.
Qian
Q
Xu
The Third Department of Surgery, Affiliated Hospital of Chengde Medical University, Chengde, Hebei 067000, P.R. China.
Enhong
E
Zhao
The Third Department of Surgery, Affiliated Hospital of Chengde Medical University, Chengde, Hebei 067000, P.R. China.
Xin
X
Zheng
Cancer Research Center, The First Affiliated Hospital of Liaoning Medical University, Jinzhou, Liaoning 121000, P.R. China.
Huachuan
H
Zheng
Cancer Research Center, The First Affiliated Hospital of Liaoning Medical University, Jinzhou, Liaoning 121000, P.R. China.
Shuang
S
Zhao
Department of Pharmacology, Chengde Medical University, Chengde, Hebei 067000, P.R. China.
Shi
S
Ding
D017209:Apoptosis / Q000187:drug effects*; D016227:Benzoquinones / Q000494:pharmacology*; D053148:Caspase 3 / Q000235:genetics / Q000378:metabolism; D053453:Caspase 9; D002453:Cell Cycle / Q000187:drug effects*; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D019938:Cyclin D1 / Q000235:genetics / Q000378:metabolism; D059565:G2 Phase Cell Cycle Checkpoints / Q000187:drug effects; D018841:HSP90 Heat-Shock Proteins / Q000037:antagonists & inhibitors; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins / Q000235:genetics / Q000378:metabolism; D047029:Lactams, Macrocyclic / Q000494:pharmacology*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D050796:STAT3 Transcription Factor / Q000235:genetics / Q000378:metabolism; D000077022:Survivin
C107489:BIRC5 protein, human; D016227:Benzoquinones; D018841:HSP90 Heat-Shock Proteins; D051034:Inhibitor of Apoptosis Proteins; D047029:Lactams, Macrocyclic; D012333:RNA, Messenger; D050796:STAT3 Transcription Factor; D000077022:Survivin; D019938:Cyclin D1; C112765:tanespimycin; D053148:Caspase 3; D053453:Caspase 9