雷公藤红素对人早幼粒细胞白血病NB4细胞增殖和凋亡的影响

摘要

Objective; To investigate the anti - proliferation and apoptosis of Celastrol on acute promyelocytic leukemia (APL) NB4 cell lines and the expression SUMO - dependent ubiquitin E3 ligase (RNF4) expression. Methods ; NB4 cells were treated with various concentrations of Celastrol, then detected the effect of Celastrol on the growth and apoptosis of NB4 cells and measured RNF4 mRNA expression level by growth curve, Cell Counting Kit - 8 assay, PI and Annexin V - FITC double staining, fluorescence microscope observation, flow cytometric analysis and real -time PCR. Results; Celastrol significantly inhibited the proliferation of NB4 cells. The 50% inhibitory concentration ( IC50 ) of Celastrol on NB4 cells was 0. 854 ± 0. 035μmol/L. NB4 cells were induced apoptosis or death by exposing to 0. 5 μmol/L and 1 μmol/L Celastrol for 48 hours. Celastrol increased RNF4 mRNA expression of NB4 cells by about 60% . Conclusion; Celastrol inhibits the growth of NB4 cells and induces cell apoptosis, which indicates Celastrol has a function of resisting APL and becomes a exploiting medicine. Furthermore, Celastrol can increase RNF4 mRNA expression of NB4 cells, which reveals Celastrol may resist APL by RNF4 related signal pathway.%目的:研究雷公藤红素对人早幼粒细胞白血病细胞系NB4的抑制作用和诱导凋亡作用以及对SU-MO依赖性特异泛素E3连接酶(RNF4)mRNA表达水平的影响.方法:用不同浓度的雷公藤红素处理NB4细胞,测定细胞生长曲线；CCK-8试剂盒检测细胞存活率；Annexin V/PI双染、荧光显微镜观察、流式细胞仪检测细胞凋亡情况；实时定量PCR检测RNF4 mRNA表达.结果:雷公藤红素能够显著抑制NB4细胞的增殖,雷公藤红素对NB4细胞处理48h的细胞半数抑制浓度为0.854±0.035 μmol/L；0.5 μmol/L和1μmol/L雷公藤红素处理NB4细胞48小时后均发生明显的细胞凋亡；0.1μmol/L雷公藤红素可使NB4细胞中的RNF4mRNA水平上调约60％.结论:雷公藤红素对急性早幼粒细胞白血病NB4细胞有明显的抗增殖和促凋亡作用,可能通过与RNF4相关的信号通路达到抗白血病作用,这为临床应用雷公藤红素治疗急性早幼粒细胞白血病提供了实验依据.