小白菊内酯联合阿霉素抑制白血病Jurkat细胞增殖的实验研究

摘要

目的：探讨小白菊内酯（ PTL）联合阿霉素（ ADM）对成人T淋巴细胞白血病Jurkat细胞增殖的影响及可能机制。方法 MTT法计算PTL和ADM 48h的半数抑制浓度（IC50），检测PTL（4、8、16μmol／L）、ADM（0�25、0�5、1μmol／L）作用于Jurkat细胞48h的增殖抑制率；流式细胞仪检测ADM（0�25、0�5、1μmol／L）、PTL（4、8、16μmol／L）以及PTL（8μmol／L）联合ADM（0�5μmol／L）作用于Jurkat细胞48h的凋亡率；免疫组化法检测ADM（0�5μmol／L）、PTL（8μmo／L）以及PTL（8μmol／L）联合ADM（0�5μmol／L）作用于Jurkat细胞48h后核因子-κBp65（ NF-κBp65）的蛋白表达。均设未加药的Jurkat细胞为对照组。结果 MTT法检测显示，PTL、ADM作用于Jurkat细胞48h的IC50分别为8�11μmol／L和0�53μmol／L。 PTL、ADM抑制Jurkat细胞的增殖呈浓度依赖性，PTL联合ADM组对Jurkat细胞的增殖抑制率高于两单药组和对照组。流式细胞术检测显示，ADM、PTL单药诱导Jurkat细胞的凋亡率随着药物浓度的增加而增高；PTL联合ADM组的细胞凋亡率为（84�50±5�64）％，显著高于两单药组和对照组（ P＜0�05）。免疫组化检测显示，单药PTL及PTL联合ADM作用于Jurkat细胞48h后NF-κBp65蛋白的阳性积分明显低于对照组（ P＜0�05），ADM单药组明显高于对照组（ P＜0�05），PTL联合ADM组均明显低于单药组（ P＜0�05）。结论 PTL可能通过抑制NF-κBp65蛋白表达，增强ADM抑制Jurkat细胞增殖能力，促进其凋亡。%Objective To investigate the inhibitory effect of parthenolide(PTL) combined with adriamycin (ADM)on prolifera-tion and apoptosis of adult acute T lymphoblastic leukemia Jurkat cells and their possible mechanism. Methods MTT assay was applied to calculate the half inhibitory concentration(IC50) of PTL and ADM for 48h, and detect the inhibition rates by treating Jurkat cells with different concentration groups of PTL(4,8,16μmol/L) or ADM (0�25,0�5,1μmol/L) for 48h. Flow cytometry was applied to detect the apoptosis rate by treating Jurkat cells with ADM (0�25, 0�5, 1μmol/L), PTL (4, 8, 16μmol/L), and PTL (8μmol/L) combined with ADM (0�5μmol/L) for 48h. Immunohistochemical method was employed to detect the expression of protein nuclear factor-κBp65 (NF-κBp65) treated by ADM (0�5μmol/L), PTL (8μmol/L), and PTL (8μmol/L) combined with ADM (0�5μmol/L). Blank control group was set for above experiments. Results MTT assay showed that PTL and ADM inhibited the growth of Jurkat cells in a dose-de-pendent manner, with IC50 of 8�11μmol/L and 0�53μmol/L at 48h. Compared with single drug groups and control group, PLT combined with ADM could significantly inhibit the proliferation of Jurkat cells. Flow cytometry showed that the apoptosis rate of Jurkat cells in PTL or ADM single agent group was in a dose-dependent manner. The apoptosis rate of combined group was (84�50±5�64) %, higher than those of single agent groups and control group ( P<0�05) . Immunohistochemical detection showed that the positive score of NF-κBp65 in PTL and PTL combined with ADM was lower than control group (P<0�05), while in ADM group was higher than control group (P<0�05), and the combined group was lower than the two single agent groups (P<0�05). Conclusion PTL may inhibit the expression of NF-κBp65 protein, raise the ability of inhibition of ADM on Jurkat cells proliferation, and promote cells apoptosis.