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目的探讨硼替佐米(Bortezomi,Bor)对急性早幼粒细胞白血病NB4细胞株和急性红白血病TF1细胞株增殖、凋亡及SALL4基因和Wnt/β-catenin信号通路下游靶基因C-myc、CCDN1表达的影响。方法分别用10、30、50 nmol/L的Bor处理NB4和TF1细胞12、24、48 h,并设对照组(不加Bor)。采用MTT法检测细胞的增殖活力;流式细胞术检测细胞凋亡率;RT-PCR和荧光定量PCR法检测细胞中SALL4、C-myc和CCDN1基因的表达。结果 Bor可抑制两种细胞的增殖,且呈时间和剂量依赖性,NB4和TF1细胞经Bor处理48 h的IC50值分别为23.97和25.36 nmol/L。Bor可诱导两种细胞凋亡,且呈剂量依赖性,经30和50 nmol/L Bor处理24 h的NB4细胞的凋亡率与对照组相比,差异有统计学意义(P<0.05);经50 nmol/L Bor处理24 h的TF1细胞的凋亡率与对照组相比,差异有统计学意义(P<0.05)。Bor可显著抑制两种细胞中SALL4基因及C-myc、CCDN1基因的表达。经50 nmol/L Bor作用24 h后,两种细胞中3种基因的表达与对照组相比,差异均有统计学意义(P<0.05)。SALL4基因与C-myc和CCDN1基因的表达明显相关(rs值分别为0.857和0.929,P均<0.01)。结论 SALL4基因及Wnt/β-catenin信号通路下游靶基因C-myc和CCDN1基因表达的抑制在Bor诱导NB4和TF1细胞凋亡的过程可能起重要作用。
Objective To investigate the effects of Bortezomi (Bor) on the proliferation and apoptosis of acute promyelocytic leukemia NB4 cell line and acute erythroleukemia TF1 cell line as well as the expression of C-myc, CCDN1 and Wnt / β-catenin signaling pathway downstream of SALL4 and Wnt / The impact of expression. Methods NB4 and TF1 cells were treated with Boron (10,30,50 nmol / L) for 12, 24 and 48 h, respectively, and control group (without Bor). Cell viability was detected by MTT assay; apoptosis rate was detected by flow cytometry; SALL4, C-myc and CCDN1 gene expression were detected by RT-PCR and real-time PCR. Results Bor inhibited the proliferation of both cells in a dose- and time-dependent manner. The IC50 values of NB4 and TF1 cells treated with Bor for 48 h were 23.97 and 25.36 nmol / L, respectively. Bor could induce both apoptosis in a dose - dependent manner. The apoptosis rate of NB4 cells treated with 30 and 50 nmol / L Bor for 24 h was significantly higher than that of the control group (P <0.05). Compared with the control group, the apoptosis rate of TF1 cells treated with 50 nmol / L Bor for 24 h was significantly different (P <0.05). Bor significantly inhibited the expression of SALL4 and C-myc and CCDN1 genes in both cells. After treated with 50 nmol / L Bor for 24 h, the expression of the three genes in the two cell lines was significantly different from that in the control group (P <0.05). The expression of SALL4 was significantly correlated with the expression of C-myc and CCDN1 (rs = 0.857 and 0.929, respectively, P <0.01). Conclusions The inhibition of the expression of C-myc and CCDN1 genes downstream of SALL4 gene and Wnt / β-catenin signaling pathway may play an important role in the apoptosis of NB4 and TF1 cells induced by Bor.