摘要
目的探讨TEOA(2α,3α,24-三羟基-12-烯-28-乌苏酸)对人急性髓系白血病(AML)细胞增殖和凋亡的影响及可能机制.方法采用CCK-8、PI染色荧光显微镜技术研究TEOA对AML细胞株Kasumi-1、KG-1α增殖、凋亡的影响;Western blot技术检测细胞抑癌蛋白P53及c-Caspase3、BaX和Cleaved-PARP的凋亡相关蛋白表达情况;流式细胞技术检测TEOA诱导白血病细胞产生ROS水平.结果CCK-8、PI染色荧光显微镜结果显示,TEOA能明显抑制AML细胞的增殖,促进细胞凋亡,其作用有浓度依赖性;Western blot结果表明,TEOA能促进凋亡相关基因P53、c-Caspase3、BaX和Cleaved-PARP的蛋白表达,Bcl-2蛋白表达降低.流式细胞术检测结果显示,当TEOA作用于急性髓系白血病细胞后,诱导细胞ROS水平增高.结论TEOA能明显抑制急性髓系白血病细胞增殖,促进其凋亡,其作用机制可能是通过增加细胞ROS水平进而激活P53凋亡通路诱导细胞凋亡.
Objective To study the ffect of TEOA(2a,3a,24-Thrihydroxyurs-12-en-28-oicacid)on the proliferation and apoptosis of acute myeloid leukemia(AML)cells and its possible mechanisms.Methods The effects of TEOA on proliferation and apoptosis of kasumi-1 and Kg-1 a of acute myeloid leukemia cell lines were studied by CCK-8 assay and PI staining fluorescence microscopy.Expression of p53 gene as well as apoptosis-associated genes c-Caspase3,BaX and Cleaved-PARP were examined by W estern blot.The ROS level of TEOA induced leukemia cells was detected by Flow cytometry.Results CCK-8 assay and PI staining fluorescence microscopys shows that TEOA significantly inhibited the proliferation of AML cells,and promote cell apoptosis,which showed concentration-dependent.Westerm blot showed that the protein expression of apoptosis-associated genes P53,c-Caspase3,BaX and Cleaved-PARP increased greatly,,and the protein expression of Bcl-2 decreased.Flow cytometry shows that the ROS levels will increase when TEOA treated the AML cells.Conclusion TEOA can inhibit the proliferation and promote the apoptosis of AML cells,which may be by increasing the levels of intracellular ROS and then activating the protein of P53 apoptotic pathways.
出处
《浙江临床医学》
2021年第5期634-637,共4页
Zhejiang Clinical Medical Journal
基金
浙江省中医药科技计划项目(2017ZA011)。