Mcl-1抑制剂UMI-77诱导胆囊癌细胞GBC-SD凋亡的机制研究-东南大学学报医学版

Mcl-1抑制剂UMI-77诱导胆囊癌细胞GBC-SD凋亡的机制研究

单位：内蒙古包钢医院肝胆外科, 内蒙古包头 014010

分类号：R735.8

出版年·卷·期（页码）：2018·37·第五期（771-776）

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、cleaved-caspase 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，10 μmol·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 10 μmol·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.

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