摘要
目的:探讨蛋白酶抑制剂依沙佐米对胰腺癌细胞凋亡和NF-κB通路的影响。方法:培养人胰腺癌细胞系CFPAC-1和PANC-1,加入浓度为0、10、20、30和40 nmol/L的依沙佐米培养12、18、24和48 h,RT-qPCR检测细胞中NF-κB p65、IκB激酶(IκB kinase, IKK)、Bax和caspase-3的mRNA表达,Western blot检测细胞中凋亡相关因子NF-κB p65、IKK、Bax和caspase3的蛋白水平,CCK-8法检测细胞活力,流式细胞术检测细胞凋亡情况。结果:CCK-8实验的检测结果显示,添加10~40 nmol/L依沙佐米均对PANC-1细胞和CFPAC-1细胞的活力具有抑制作用(P<0.05),呈时间和剂量依赖性;细胞凋亡检测结果显示随着依沙佐米浓度的增加,PANC-1细胞和CFPAC-1细胞的凋亡率均显著增加(P<0.05);与添加0 nmol/L抑制剂的对照细胞比较添加依沙佐米后PANC-1细胞和CFPAC-1细胞NF-κB p65和IKK的mRNA表达水平显著降低(P<0.05);凋亡因子Bax和caspase-3的表达水平均显著升高(P<0.05)。添加依沙佐米后PANC-1细胞和CFPAC-1细胞的NF-κB p65和IKK的蛋白水平均显著降低(P<0.05),与mRNA检测结果一致;CFPAC-1细胞的凋亡因子Bax和caspase-3蛋白表达水平均显著升高(P<0.05),PANC-1细胞的caspase-3蛋白表达水平显著升高,Bax蛋白在依沙佐米为10 nmol/L浓度时变化的差异无统计学显著性。结论:蛋白酶体抑制剂依沙佐米可能通过抑制NF-κB通路的激活抑制胰腺癌细胞的活力,促进细胞凋亡,且作用效果呈时间和剂量依赖性。
AIM: To investigate the effects of ixazomib on the apoptosis and NF-κB signaling pathway in pancreatic cancer cells. METHODS: Human pancreatic cancer cell lines CFPAC-1 and PANC-1 were cultured, and the cells were treated with ixazomib at 0, 10, 20, 30 and 40 nmol/L for 12, 18, 24 and 48 h. The expression of NF-κB p65, IκB kinase(IKK), Bax and caspase-3 in the cells at mRNA and protein levels was determined by RT-qPCR and Western blot. The cell viability was measured by CCK-8 assay. The apoptosis was analyzed by flow cytometry. RESULTS: Treatment with ixazomib at 10~40 nmol/L inhibited the viability of PANC-1 cells and CFPAC-1 cells, and the inhibitory rate was increased significantly with the increases in the concentration and time(P<0.05). Compared with the control cells, treatment with ixazomib significantly increased the apoptotic rates of PANC-1 cells and CFPAC-1 cells in a dose-dependent manner(P<0.05), and significantly decreased the mRNA expression levels of NF-κB p65 and IKK in the PANC-1 cells and CFPAC-1 cells(P<0.05), while the mRNA expression levels of apoptotic factors Bax and caspase-3 in the PANC-1 cells and CFPAC-1 cells were significantly increased(P<0.05). The results of Western blot showed that treatment with ixazomib significantly decreased the protein levels of NF-κB p65 and IKK in the PANC-1 cells and CFPAC-1 cells(P<0.05), which was consistent with the results of mRNA expression. The protein levels of apoptosis factors Bax and caspase-3 in the CFPAC-1 cells were significantly increased(P<0.05), and the protein level of caspase-3 in the PANC-1 cells was increased significantly(P<0.05). However, Bax protein did not increase significantly in 10 nmol/L ixazomib group. CONCLUSION: Ixazomib, a proteasome inhibitor, inhibits the viability of pancreatic cancer cells and promotes apoptosis by inhibiting the activation of NF-κB signaling pathway in a time-and dose-dependent manner.
作者
王自闯
陈小永
张娟
WANG Zi-chuang;CHEN Xiao-yong;ZHANG Juan(Henan University of Traditional Chinese Medicine, Zhengzhou 450002, China)
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2019年第8期1416-1422,共7页
Chinese Journal of Pathophysiology
基金
国家自然科学基金资助项目(No.81303096)