Xu Zheng, Song Ling, Wu Yu-Hui, Cao Bo
Department of Hematology, Xinyang Central Hospital, Xinyang 464000, Henan Province， China,E-mail:
Department of Hematology, Xinyang Central Hospital, Xinyang 464000, Henan Province， China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2022 Jun;30(3):695-703. doi: 10.19746/j.cnki.issn.1009-2137.2022.03.006.
AbstractObjective: To explore the effect and mechanism of curcumin on human T-cell acute lymphoblastic leukemia (T-ALL) cell apoptosis induced by Mcl-1 small molecule inhibitors UMI-77.
T-ALL cell line Molt-4 was cultured, and the cells were treated with different concentrations of curcumin and Mcl-1 small molecule inhibitor UMI-77 for 24 h. The MTT method was used to detect the cell survival rate after different treatment; According to the results of curcumin and UMI-77, the experimental settings were divided into control group, curcumin group (20 μmol/L curcumin treated cells), UMI-77 group (15 μmol/L Mcl-1 small molecule inhibitor UMI-77 treated cells) and curcumin+ UMI-77 group (20 μmol/L curcumin and 15 μmol/L Mcl-1 small molecule inhibitor UMI-77 treated cells), MTT method was used to detect cell proliferation inhibition rate, Annexin V-FITC/PI double staining method and TUNEL staining were used to detect cell apoptosis, DCFH-DA probe was used to detect cell reactive oxygen species, JC-1 fluorescent probe was used to detect mitochondrial membrane potential, Western blot was used to detect the expression levels of apoptosis-related proteins and Notch1 signaling pathway-related proteins.
After the treatment of Molt-4 cells with different concentrations of curcumin and Mcl-1 small molecule inhibitor UMI-77, the cell survival rate was decreased (P<0.05); Compared with the control group, the cell proliferation inhibition rate of the curcumin group and the UMI-77 group were increased, the apoptosis rate of cell was increased, the level of ROS was increased, the protein expression of Bax, Caspase-3 and Caspase-9 in the cells were all increased, and the protein expression of Bcl-2 was reduced (P<0.05); Compared with the curcumin group or UMI-77 group, the cell proliferation inhibition rate and apoptosis rate of the curcumin+UMI-77 group were further increased, and the level of ROS was increased. At the same time, the protein expression of Bax, Caspase-3 and Caspase-9 in the cells were all increased, the protein expression of Bcl-2 was reduced (P<0.05); In addition, the mitochondrial membrane potential of the cells after curcumin treatment was decreased, and the proteins expression of Notch1 and HES1 were reduced (P<0.05).
Curcumin can enhance the apoptosis of T-ALL cells induced by Mcl-1 small molecule inhibitor UMI-77 by reducing the mitochondrial membrane potential, the mechanism may be related to the inhibition of Notch1 signaling pathway.
摘要目的：探讨姜黄素对Mcl-1小分子抑制剂UMI-77诱导的人T细胞急性淋巴细胞白血病（T-ALL）细胞凋亡的影响及机制。
培养T-ALL细胞系Molt-4，用不同浓度的姜黄素和Mcl-1小分子抑制剂UMI-77处理细胞24小时。采用MTT法检测不同处理后的细胞存活率；根据姜黄素和UMI-77的结果，将实验设置分为对照组、姜黄素组（用20μmol/L姜黄素处理的细胞）、UMI-77组（用15μmol/L Mcl-1小分子抑制剂UMI-77处理的细胞）和姜黄素+UMI-77组（用20μmol/L姜黄素和15μmol/L Mcl-1小分子抑制剂UMI-77处理的细胞），采用MTT法检测细胞增殖抑制率，采用Annexin V-FITC/PI双染法和TUNEL染色法检测细胞凋亡，采用DCFH-DA探针检测细胞活性氧，采用JC-1荧光探针检测线粒体膜电位，采用Western blot检测凋亡相关蛋白和Notch1信号通路相关蛋白的表达水平。
用不同浓度的姜黄素和Mcl-1小分子抑制剂UMI-77处理Molt-4细胞后，细胞存活率降低（P<0.05）；与对照组相比，姜黄素组和UMI-77组的细胞增殖抑制率升高，细胞凋亡率升高，活性氧水平升高，细胞内Bax、Caspase-3和Caspase-9的蛋白表达均升高，Bcl-2的蛋白表达降低（P<0.05）；与姜黄素组或UMI-77组相比，姜黄素+UMI-77组的细胞增殖抑制率和凋亡率进一步升高，活性氧水平升高。同时，细胞内Bax、Caspase-3和Caspase-9的蛋白表达均升高，Bcl-2的蛋白表达降低（P<0.05）；此外，姜黄素处理后细胞的线粒体膜电位降低，Notch1和HES1的蛋白表达降低（P<0.05）。
姜黄素可通过降低线粒体膜电位增强Mcl-1小分子抑制剂UMI-77诱导的T-ALL细胞凋亡，其机制可能与抑制Notch1信号通路有关。
