Mcl-1抑制剂UMI-77诱导胆囊癌细胞GBC-SD凋亡的机制研究  

Mechanism of apoptosis induced by Mcl-1 inhibitor UMI-77 on gallbladder carcinoma GBC-SD cells

作　　者：张生彬[1] 刘宝琴[1] 董长城[1] 李冰[1] ZHANG Shengbin;LIU Baoqin;DONG Changcheng;LI Bing(Department of Hepatobiliary Surgery,BaoGang Hospital,Baotou 014010,China)

机构地区：[1]内蒙古包钢医院肝胆外科,内蒙古包头014010

出　　处：《东南大学学报（医学版）》2018年第5期771-776,共6页Journal of Southeast University(Medical Science Edition)

基　　金：内蒙古自治区自然科学基金资助项目(2013MS1102)

摘　　要：目的:探讨髓样细胞白血病-1(Mcl-1)抑制剂UMI-77诱导胆囊癌细胞GBC-SD凋亡的机制。方法:采用不同浓度的UMI-77处理胆囊癌GBC-SD细胞,MTT比色法和Annexin V/PI分别检测细胞增殖和细胞凋亡情况; Western blotting检测UMI-77作用后细胞中Mcl-1、Bcl-2、Bcl-xL、Bax、Bak、cleaved-caspase 9、cleavedcaspase 3和cleaved-PARP蛋白的表达。结果:MTT检测结果显示,不同浓度的UMI-77分别作用于GBC-SD细胞后均对细胞增殖产生不同程度的抑制作用,且呈剂量依赖性和时间依赖性。Annexin V/PI检测结果显示,随着UMI-77作用浓度的升高,细胞凋亡率逐渐增加,且呈剂量依赖性。Western blotting结果显示,10μmol·L^(-1)UMI-77处理GBC-SD细胞24 h后,其抗凋亡蛋白Mcl-1的表达量显著降低(P <0. 05),促凋亡蛋白Bax和Bak的表达量显著升高(P <0. 05),但Bcl-2和Bcl-xL的蛋白表达量无显著性变化,且其cleaved-caspase 9、cleaved-caspase 3和cleaved-PARP蛋白的表达量均显著升高(P <0. 05)。结论:Mcl-1抑制剂UMI-77可通过caspase介导的内源性凋亡途径诱导胆囊癌GBC-SD细胞发生凋亡,且呈剂量依赖性,因此,Mcl-1可能成为胆囊癌研究中一个新的治疗靶点。Objective:To investigate the mechanism of apoptosis induced by myeloid cell leukemia-1(Mcl-1)inhibitor UMI-77 on gallbladder carcinoma GBC-SD cells.Methods:GBC-SD cells were treated with different concentrations of UMI-77.The proliferation and apoptosis of GBC-SD cells were detected by MTT assay and Annexin V/PI.The expressions of Mcl-1,Bcl-2 Bcl-xL,Bax,Bak,leaved-caspase 9,cleaved-caspase 3 and cleaved-PARP protein in GBC-SD cells after treated with UMI-77 were detected by western blotting.Results:The results of MTT showed that different concentrations of UMI-77 had different inhibitory effects on cell proliferation of GBC-SD cells in a dose-dependent and time-dependent manner.Annexin V/PI test results showed that the apoptotic rate increased gradually with the increase of UMI-77 concentration in a dose-dependent manner.Western blotting showed that the expression of anti-apoptotic protein Mcl-1 was significantly decreased(P<0.05),and the expressions of Bax and Bak proteins were significantly increased(P<0.05),but the expressions of Bcl-2 and Bcl-xL proteins had no statistical significance,and the expression levels of cleaved-caspase 9,cleaved-caspase 3 and cleaved-PARP proteins were significantly increased(P<0.05)after treatment with 10μmol·L-1 UMI-77 for 24 h.Conclusion:Mcl-1 inhibitor UMI-77 can induce apoptosis of GBC-SD cells in a dose-dependent manner by caspase-mediated endogenous apoptosis pathway,therefore,Mcl-1 may become a new therapeutic target in the gallbladder cancer research.

关 键 词：Mcl-1抑制剂 UMI-77 胆囊癌 细胞凋亡 

分 类 号：R735.8[医药卫生—肿瘤]