柔红霉素诱导急性淋巴细胞白血病Jurkat细胞凋亡的作用因素研究

目的 探讨柔红霉素(DNR)致急性淋巴细胞白血病Jurkat细胞株凋亡的作用因素.方法 不同浓度(0、0.05、0.1、0.5、1μg/mL)DNR及N-乙酰半胱氨酸(NAC)预处理组作用于Jurkat细胞株,采用MTT法检测细胞活力,Hoechst/PI双染法观察细胞凋亡,流式细胞术检测细胞活性氧(ROS)浓度及凋亡,RT-PCR法检测bax、survivin、bcl-2和bcl-xl基因的表达.结果 DNR可明显抑制Jurkat细胞株的活性,0、0.05、0.1、0.5、μg/mL的DNR及NAC预处理组分别作用于Jurkat细胞株24 h后,ROS水平分别为(7.98±0.55)%、(8.88±0.86)%、(9.46±0.98)%、(17.48±2.98)%、(24.46±2.43)%和(11.59±1.29)%,加入NAC后ROS生成受到抑制(P<0.01);细胞凋亡率分别为(11.41±1.44)%、(34.96±3.32)%、(45.58±3.12)%、(84.19±2.65)%、(87.93±1.74)%和(80.47±0.63)%,NAC预处理组与0.5 μg/mL DNR组比较,细胞凋亡率变化无统计学意义(P>0.05).NAC抑制bax基因mRNA表达,上调survivin、bcl-2和bcl-xl表达(均P<0.05).结论 DNR能够诱导Jurkat细胞株凋亡;ROS参与了DNR诱导的Jurkat细胞株凋亡;凋亡相关基因bax、bcl-2、bcl-xl及survivin能够与ROS相互作用,调节DNR诱导的Jurkat细胞株凋亡.

Influencing factors of daunorubicin induced apoptosis in Jurkat cells of acute lymphoblastic leukemia

Objective To explore the influencing factors of daunorubicin (DNR) induced apoptosis in Jurkat cells of acute lymphoblastic leukemia. Methods Jurkat cells were treated with different concentrations of DNR(0, 0.05, 0.1, 0.5 and 1 μg/mL)or pretreated with N-acetylcysteine (NAC). Cell proliferation was detected by MTT assay, cell apoptosis was observed by Hoechst/PI, reactive oxygen species (ROS) level and apoptosis were determined by flow cytometry, and the expression of survivin, bax, bcl-2 and bcl-xl mRNA was examined by RT-PCR. Results DNR siguificantly inhibited the proliferation of Jurkat cells. After treatment with 0, 0.05, 0.1, 0.5, 1 μg/mL DNR and pretreated with NAC for 24 h, ROS levels were (7.98±0.55)%, (8.88±0.86)%, (9.46±0.98)%, (17.48±2.98)%, (24.46±2.43)% and (11.59±1.29)%, respectively, and cell apoptosis rates were (11.41±1.44)%, (34.96±3.32)%, (45.58± 3.12)%, (84.19±2.65)%, (87.93±1.74)% and (80.47±0.63)%, respectively. After pretreatment with NAC, the levels of ROS were siguifieantly inhibited (P < 0.01). There was no significant difference in apoptosis rates between treatment with 0.5μg/mL DNR and pretreatment with NAC(P>0.05). The expression of bax mRNA was down regulated and the expression of survivin, bcl-2 and bcl-xl was up regulated (P < 0.05). Conclusion DNR can induce apoptosis of Jurkat cells. ROS may participate in the DNR induced Jurkat cell apoptosis, and apoptosis-related gene bax, bcl-2, bcl-xl and survivin may interact with ROS in the regulation of DNR induced Jurkat cell apoptosis.