摘要
目的探讨硼替佐米(BOR)对急性T淋巴细胞白血病细胞株Jurkat的增殖抑制和促凋亡作用及其相关作用机制。方法2017年5月至2019年5月,采用四甲基偶氮唑蓝法(MTT法)检测硼替佐米对Jurkat细胞增殖的影响;流式细胞仪检测硼替佐米对Jurkat细胞细胞凋亡的影响;实时定量聚合酶链反应(RT-PCR)法检测硼替佐米对Jurkat细胞Bax、Bcl-2、Cox-2基因表达水平的影响。结果5 ng/mL、10 ng/mL、20 ng/mL、40 ng/mL浓度BOR作用于Jurkat细胞24 h的增殖抑制率分别为(13.23±0.71)%、(39.53±0.95)%、(53.07±1.12)%、(60.43±0.75)%,48 h的增殖抑制率分别为(25.20±0.96)%、(52.80±1.30)%、(60.67±0.64)%、(75.10±1.35)%,72 h的增殖抑制率分别为(38.37±0.93)%、(60.94±0.85)%、(73.83±5.08)%、(88.37±1.55)%,Jurkat细胞的增殖抑制率随药物浓度的增加、作用时间的延长而增加,差异均有统计学意义(F=1602.202、1085.089、181.034,均P<0.05)。5 ng/mL、10 ng/mL、20 ng/mL、40 ng/mL浓度BOR作用于Jurkat细胞24 h、48 h、72 h,Jurkat细胞凋亡率随药物浓度的增加、作用时间的延长而增加,差异均有统计学意义(F=1288.571、223.378、251.175,均P<0.05)。5 ng/mL、10 ng/mL、20 ng/mL、40 ng/mL浓度BOR作用于Jurkat细胞24 h、48 h、72 h,Jurkat细胞Bax mRNA表达水平随药物浓度的增加、作用时间的延长而增高,差异均有统计学意义(F=258.446、518.929、276.764,均P<0.05);而Bcl-2 mRNA、Cox-2 mRNA表达水平随药物浓度的增加、作用时间的延长而减低,差异有统计学意义(FBcl-2 mRNA=236.848、264.849、343.968,FCox-2 mRNA=679.404、1288.681、1541.850,均P<0.05)。结论BOR对Jurkat细胞有抑制增殖、诱导凋亡的作用。BOR可使Jurkat细胞BaxmRNA表达水平上调、Bcl-2和Cox-2 mRNA表达水平下调。BOR上调Bax表达、下调Bcl-2和Cox-2表达是其促凋亡的分子机制之一。
Objective To explore the effects and mechanisms of bortezomib on the proliferation and apoptosis of acute T lymphocyte leukemia cell line Jurkat.Methods MTT assay was used to test the influence of bortezomib on the proliferation of Jurkat cells.Flow cytometry was used to detect the influence of bortezomib on apoptosis of Jurkat cells.Real-time quantitative polymerase reaction(RT-PCR)was used to detect the effects of bortezomib on the expression of Bax,Bcl-2 and Cox-2 genes in Jurkat cells.Results The inhibition rates of 5ng/mL,10ng/mL,20ng/mL and 40ng/mL bortezomib on Jurkat cells at 24h were(13.23±0.71)%,(39.53±0.95)%,(53.07±1.12)%,(60.43±0.75)%,respectively,and the inhibition rates at 48h were(25.20±0.96)%,(52.80±1.30)%,(60.67±0.64)%,(75.10±1.35)%,respectively.The inhibitory rates of proliferation of Jurkat cells at 72h were(38.37±0.93)%,(60.94±0.85)%,(73.83±5.08)%,(88.37±1.55)%,respectively.The inhibitory rates of proliferation of Jurkat cells increased with the increase of drug concentration and the prolongation of action time,and the differences were statistically significant(F=1602.202,1085.089,181.034,all P<0.05).Bortezomib(5ng/mL,10ng/mL,20ng/mL and 40ng/mL)treatment for 24h,48h and 72h,the apoptosis rate of Jurkat cells increased with the increase of drug concentration and the prolongation of action time,the differences were statistically significant(F=1288.571,223.378,251.175,all P<0.05).The expression of Bax mRNA in Jurkat cells increased with the increase of drug concentration and time(F=258.446,518.929,276.764,all P<0.05).The Bcl-2 mRNA and Cox-2 mRNA expression levels decreased with the increase of drug concentration and the prolongation of action time(F Bcl-2 mRNA=236.848,264.849,343.968,F Cox-2 mRNA=679.404,1288.681,1541.850,all P<0.05).Conclusion Bortezomib can inhibit the proliferation and induce apoptosis of Jurkat cells.Bortezomib can increase the expression of Bax mRNA and decrease the expression of Bcl-2 and Cox-2 mRNA,which may be the molecular mechanism of bortezomib to promote apoptosis.
作者
葛文军
马梁明
曹丽萍
田雪娇
杨晶
冯芳
陈玉芬
孙涛
Ge Wenjun;Ma Liangming;Cao Liping;Tian Xuejiao;Yang Jing;Feng Fang;Chen Yufen;Sun Tao(Department of Hematology,the Fifth People's Hospital of Datong,Datong,Shanxi 037000,China)
出处
《中国基层医药》
CAS
2020年第14期1693-1697,共5页
Chinese Journal of Primary Medicine and Pharmacy
