Molecular mechanisms involved in curcumol induced apoptosis in human gastric carcinoma cell line BGC823
Hong Zhang, Haijun Jiang, Jianguo Gao, Jian-qing Du, Zhi Yang
Abstract
Hong Zhang, Haijun Jiang, Jianguo Gao, Jian-qing Du, Zhi Yang
Abstract
Objective To investigate the effect of curcumol on apoptosis of human gastric carcinoma cell line BGC823 and the molecular nechanisms.Methods BGC823 cells were cultured and treated with different curcumol concentration (12.5,25,50 and 100 mg/L) for 24 h and 48 h,and the growth inhibition were tested by thiazolyl blue terazolium bromide (MTF) assay.Flow cytometry (FCM) were used to measure the cell apoptosis rate and cell cycle of BGC823 cells.Caspase-3 activity was assessed by colorimetric assay.Cells treated with 100 mg/L curcunol for 48 h were collected and subjected to RTPCR and Western blot assays for the expression of Caspase-3,Bcl-2,Bax and Survivin.Results There was a time-and dose-dependent inhibition of cell proliferation of BGC823 cells by curcumol.Tbe cells in G0/G1 phase increased,and in S phase decreased on exposure to curcumol for 24 h.FCM analysis also indicated that the apoptosis rate of BGC823 cells increased in dose-dependent manner (P < 0.05).Curcumol increased the activity of Caspase-3 dose-dependently (P < 0.05).RT-PCR and Western blot indicated that curcumol decreased Bcl-2 and Survivin expression as well as increased Caspase-3 and Bax expression (P < 0.05).Conclusions Curcumol inhibits BGC823 cell growth,arresting cells in G0/G1 phase and inducing cell apoptosis.The mechanism may be related with increasing the activity of Caspase-3,down-regulating the expression of Bcl-2 and Survivin,and up-regulating the expression of Caspase-3 and Bax. Key words: Stomach neoplasms ; Apoptosis ; Cell proliferation; Cell cycle
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Objective To investigate the effect of curcumol on apoptosis of human gastric carcinoma cell line BGC823 and the molecular nechanisms.Methods BGC823 cells were cultured and treated with different curcumol concentration (12.5,25,50 and 100 mg/L) for 24 h and 48 h,and the growth inhibition were tested by thiazolyl blue terazolium bromide (MTF) assay.Flow cytometry (FCM) were used to measure the cell apoptosis rate and cell cycle of BGC823 cells.Caspase-3 activity was assessed by colorimetric assay.Cells treated with 100 mg/L curcunol for 48 h were collected and subjected to RTPCR and Western blot assays for the expression of Caspase-3,Bcl-2,Bax and Survivin.Results There was a time-and dose-dependent inhibition of cell proliferation of BGC823 cells by curcumol.Tbe cells in G0/G1 phase increased,and in S phase decreased on exposure to curcumol for 24 h.FCM analysis also indicated that the apoptosis rate of BGC823 cells increased in dose-dependent manner (P < 0.05).Curcumol increased the activity of Caspase-3 dose-dependently (P < 0.05).RT-PCR and Western blot indicated that curcumol decreased Bcl-2 and Survivin expression as well as increased Caspase-3 and Bax expression (P < 0.05).Conclusions Curcumol inhibits BGC823 cell growth,arresting cells in G0/G1 phase and inducing cell apoptosis.The mechanism may be related with increasing the activity of Caspase-3,down-regulating the expression of Bcl-2 and Survivin,and up-regulating the expression of Caspase-3 and Bax. Key words: Stomach neoplasms ; Apoptosis ; Cell proliferation; Cell cycle
Key concepts: Survivin, Apoptosis, Western blot, Molecular biology, Cell culture, Cell cycle, Flow cytometry, Cell