硼替佐米联合氟达拉滨对淋巴瘤细胞系HUT-78的体外杀伤效应

Cytotoxic Effects of Bortezomib Combined with Fludarabine in T-cell Lymphoma Cell Line HUT-78 In Vitro

目的:探讨硼替佐米(Bortezomib,PS-341,BTZ)联合氟达拉滨(Fludarabine,FLU)不同给药顺序对于T细胞淋巴瘤细胞HUT-78增殖和凋亡的影响。方法:MTT法测定BTZ、FLU单药作用于HUT-78细胞系的IC_(50)。流式细胞术检测BTZ、FLU单药对HUT-78细胞周期分布的影响,以及两者不同顺序给药时细胞凋亡的情况。Western blot检测不同给药顺序作用于HUT-78细胞后cleaved caspase-3的表达情况。结果:BTZ、FLU单药作用于HUT-78细胞24 h的IC_(50)分别是(53.84±9.31)nmol/L和(2.54±0.33)mmol/L。细胞周期分析显示BTZ主要使细胞停滞在S期,而FLU主要使细胞停滞在G_0～G_1期;而在细胞凋亡方面BTZ→FLU组促使细胞凋亡的作用与同时给药及FLU→BTZ组相比最为显著(P<0.01)。同样,Western blot结果显示BTZ→FLU组cleaved caspase-3表达水平最高(P<0.01)。结论:BTZ与FLU联合使用可有效抑制HUT-78细胞的生长,先用BTZ再用FLU的给药顺序的促凋亡作用最强,该用药顺序可为临床用药所借鉴。

Objective：To evaluate the schedule-dependent effect of bortezomib （BTZ） combined with fludarabine （FLU） on the cell proliferation and apoptosis of the HUT-78cell line. Methods：The half-maximal inhibitory concentration （IC50） of BTZ and FLU was estimated by3-（ 4,5-deimethylthiazol- 2-yl）- 2,5-diphenyltetrazolium bromide. Flow cytometry was used to analyze the effects of BTZ or FLU alone on the cell cycle, as well as the rate of apoptosis when they were combined in different sequences. The protein level of cleaved caspase- 3 in HUT- 78cells was determined by Western blot. Results： The IC 50of BTZ or FLU alone for HUT- 78cell were 53.84nmol/L ± 9.31nmol/L and 2.54mmol/L ±0.33mmol/L, respectively, at 24h. BTZ promoted cell cycle arrest at the S phase, whereas FLU blocked cells at the G0 –G1 phase. In cell apoptosis, the rate of the BTZ→FLU group was higher than those of other groups （P〈0.01）. Western blot showed that compared with the concurrent group and the FLU →BTZ group, the protein level of cleaved caspase-3 was highest in the BTZ→FLU group （P〈0.01）. Conclusion：BTZ combined with FLU, especially BTZ followed by FLU, in-duces a higher apoptosis rate in the HUT- 78cell line. This sequential usage of BTZ and FLU is available as a clinical therapy.