南瓜蛋白对人胰腺癌细胞CFPAC-1增殖抑制及诱导凋亡作用

Proliferation Inhibition and Apoptosis Induction of Cucurmosin on Human Pancreatic Cancer CFPAC-1 Cells

目的探讨南瓜蛋白(CUS)体外对人胰腺癌细胞CFPAC-1的增殖抑制及诱导凋亡作用。方法甲基噻唑基四唑法检测南瓜蛋白对CFPAC-1细胞的增殖抑制作用,透射电镜观察细胞凋亡的形态学变化,流式细胞仪检测细胞周期及凋亡率,蛋白质印迹法检测caspase-3及bcl-2蛋白表达水平。结果不同浓度(0.031 25、0.062 5、0.125、0.25、0.5、1及2μmol.L-1)南瓜蛋白作用不同时间(24、48及72 h)后,南瓜蛋白对胰腺癌细胞有增殖抑制作用,并呈时间和剂量依赖性(P<0.05)。1μmol.L-1南瓜蛋白作用72 h后,电镜下可见明显凋亡改变,并可见凋亡小体。流式细胞仪结果示,与对照组相比,S期细胞减少,G0/G1期细胞增多,细胞由G0/G1期进入S期延缓;不同浓度(0、0.062 5、0.25及1μmol.L-1)南瓜蛋白作用72 h后细胞凋亡率分别为(0.33±0.37)%、(19.26±1.49)%、(37.13±2.07)%及(55.64±2.91)%。随着药物浓度的增高,caspase-3蛋白表达逐渐增强,bcl-2蛋白表达逐渐减弱。结论南瓜蛋白对人胰腺癌细胞CFPAC-1具有明显抑制作用;南瓜蛋白可能通过上调caspase-3及下调bcl-2蛋白表达诱导胰腺癌细胞凋亡。

BJECTIVE To investigate the mechanism that cucurmosin（CUS） induces the apoptosis of human pancreatic cancer cell line CFPAC-1. METHODS The inhibition effect of cucurmosin on CFPAC-1 cell was detected by MTT assay.The apoptosis was observed by transmission electron microscope.The cell cycle and apoptosis rate were analyzed by flow cytometry.The expressions of caspase-3 and bcl-2 protein were determined by Westernblot. RESULTS After CFPAC-1 cells were treated with cucurmosin of 0.031 25,0.062 5,0.125,0.25,0.5,1 and 2 μmol·L-1 for 24,48 and 72 h,the proliferation of CFPAC-1 cells was inhibited in a time-and dose-dependent manner（P〈0.05）.After CFPAC-1 cells were treated with 1 μmol·L-1 cucurmosin for 72 h,typical apoptosis changes were observed under transmission electron microscope.Compared with control group,more cells were arrested at G0/G1 phases（P〈0.05） and fewer cells were at S phases（P〈0.05）.CUS decreased the speed of cell-cycle progression from G0/G1 phase into S phase.After CFPAC-1 cells were treated without（control） or with cucurmosin of 0.062 5,0.25 and 1 μmol·L-1 for 72 h,the apoptosis rates of CFPAC-1 cells were（0.33±0.37）%,（19.26±1.49）%,（37.13±2.07）% and（55.64±2.91）%,respectively.The expression of caspase-3 was elevated,whereas the expression of Bcl-2 was lessened gradually. CONCLUSION Cucurmosin induces the apoptosis of pancreatic cancer CFPAC-1 cells through up-regulating the expression of caspase-3 and down-regulating the expression of bcl-2.