【摘要】 目的:探讨小剂量丙戊酸钠(valproate acid,VPA)对吡柔比星(pirarubicin,THP)抑制儿童急性B淋巴细胞白血病(B-cell acute lymphoblastic leukemia,B-ALL)细胞株Raji细胞生长的增敏作用及其可能机制。方法:应用WST-1法检测不同浓度的VPA、THP单药及二者联合应用时对Raji细胞增殖的影响;将筛选出最佳联合作用浓度的THP与VPA联合作用于Raji细胞,显微镜下观察细胞的增殖情况,TUNEL法检测细胞凋亡,同时通过Real-time PCR、Western Blot检测不同组合药物作用前后细胞凋亡相关蛋白Bax、Bcl-2 mRNA和蛋白水平的表达变化。结果:不同浓度VPA(0.25、0.5、1、2、3、4、5 mmol/L)、THP(0.1、1.0、2.5、5、10μmol/L)对Raji细胞均有一定的增殖抑制作用,呈时间和剂量依赖性;联合用药对细胞的增殖抑制率明显高于THP单药组,也呈时间和剂量依赖性(P<0.05)。联合用药组细胞凋亡率明显高于VPA和THP单药组。Real-time PCR、Western Blot结果显示联合用药组(0.25 mmol/L VPA+2.5μmol/L THP)Bax mRNA和蛋白表达水平显著高于单药组,Bcl-2 mRNA和蛋白表达量显著低于单药组,各组和对照组相比差异均有统计学意义(P<0.05)。结论:VPA及THP对Raji细胞增殖有抑制作用,且有浓度和时间依赖性。低浓度的VPA联合不同浓度THP对Raji细胞增殖抑制作用较单药应用显著增强。VPA联合THP一定程度上通过促进凋亡进而抑制了Raji细胞的增殖作用。更多还原

【Abstract】 Objective: To study the effects of low dose of sodium valproate(valproate acid,VPA) of pirarubicin(pirarubicin,THP)inhibition in childhood B-cell acute lymphoblastic leukemia(B-ALL) cell line Raji and investigate their sensitization effect of cell growth and its possible mechanism.Methods: WST-1 was used to detect different concentration of VPA,THP and their joint application of Raji cell proliferation.The optimum concentration of THP and VPA were combined on Real-time cells.Observed the proliferation of cells under the microscope; apoptosis was detected by TNUEL; at the same time,Realtime PCR and Western Blot were used to detect the expression of Bax,Bcl-2 mRNA and protein before and after treated with different combinations of drugs.Results: VPA(0.25,0.5,1,2,3,4,5 mmol/L) and THP(0.1,1,2.5,5,10 μmol/L)had an inhibitory effect on Raji cells was in a time and dose dependent respectively.The inhibition rate of cell proliferation in combination group was significantly higher than that of THP single drug group,also in a time and dose dependent(P<0.05).Cell apoptosis rate of combination group was obviously higher than that of control group,also higher than that of VPA and THP single drug group.Real-time PCR and Western blot results showed that Bax mRNA and protein expression levels in the combined treatment group(0.25 mmol/L+2.5 mol/L THP) were significantly higher than the single drug group,Bax mRNA and protein expression were significantly lower than drug groups,while Bcl-2 mRNA and protein expression were significantly higher than drug groups,each group differences were statistical significant(P<0.05).Conclusions: VPA and THP had inhibitory effect on the proliferation of Raji cells,and there were a time and concentration dependent.The inhibition rate of Raji cells proliferation in VPA combined with different concentration of low concentration of THP was significantly more enhanced compared with single drug group.VPA combined with THP inhibited the proliferation of Raji cells,to some extent,by promoting apoptosis.更多还原