Brucine in inducing cell apoptosis of human pancreatic cancer CFPAC-1 cells by regulating the pathway of mitochondrial apoptosis / 中华胰腺病杂志

ABSTRACT

Objective To investigate the influence of Brucine on cell apoptosis of pancreatic cancer CFPAC-1 cells and the possible mechanism. Methods Brucine in different concentrations were used to treat CFPAC-1 cells. Cell proliferation was determined by MTT assay and cell apoptosis was determined by flow cytometer assay. Mitochondrial membrane potential was examined by JC-1 staining. The protein expression of Bax and Bcl-2 was measured by Western Blot. Results The growth inhibition rates of CFPAC-1 cells after being treated with 0 (control group), 0. 4 and 0. 8 mmol/L Brucine for 24, 48 and 72 h were 0,(30. 23 ± 0. 55)％,( 40. 61 ± 0. 15 )％, ( 46. 98 ± 1. 27 )％ and ( 50. 17 ± 0. 75 )％, ( 61. 23 ± 0. 91 )％, ( 70. 32 ± 0. 40)％, increasing with a concentration-and time-dependent increase, which was higher than that in control group; and the differences between either two groups at different time points were statistically significant (P<0. 05). CFPAC-1 cell apoptosis rate after being treated with 0, 0. 4 and 0. 8 mmol/L Brucine for 48 h was ( 2. 92 ± 0. 46 )％, ( 4. 64 ± 1. 31 )％ and ( 13. 09 ± 0. 65 )％, which increased gradually with the increased drug concentration. The apoptotic rate in 0. 8 mmol/L treatment group was obviously higher than that in control group, and the difference was statistically significant (P<0.05). With the increase of the drug concentration, the red fluorescence gradually decreased, and the green fluorescence gradually increased, indicating that the mitochondrial membrane potential was severely damaged and thus decreased. The protein expression of Bcl-2 in CFPAC-1 cells were(0.92 ±0.12),(0.67 ±0.14)and(0.35 ±0.14)mmol/L,and the expression of Bax in CFPAC-1 cells were(0. 56 ± 0. 12),(0. 85 ± 0. 10)and(1. 15 ± 0. 12)mmol/L. With the increase of brucine concentration, the expression of Bcl-2 was significantly reduced while the expression of Bax was significantly increased;and the difference was statistically significant (P<0. 05). Conclusions Brucine can effectively induce the apoptosis of human pancreatic cancer CFPAC-1 cells through mitochondrial apoptotic pathway by up-regulating the expression of Bax and down-regulating the expression of Bcl-2.