川芎嗪联合三氧化二砷对人急性早幼粒细胞白血病HL-60细胞增殖的影响

目的 探讨川芎嗪(TMP)联合三氧化二砷(As2O3)对人急性早幼粒细胞白血病(APL)细胞株HL-60增殖的影响及其治疗APL的作用机制.方法 采用HL-60和脐带内皮细胞系ECV-304两种细胞株,分为空白组、TMP组、As2O3组、TMP+ As2O3组,每组设4个复孔,经TMP单独或联合As2O3作用后,应用噻唑蓝(MTT)实验测定HL-60细胞增殖变化；检测HL-60细胞血管内皮生长因子(VEGF) mRNA和蛋白表达；台盼蓝染色计数绘制ECV-304细胞生长曲线；流式细胞仪检测ECV-304细胞早期凋亡率；倒置显微镜观察ECV-304细胞贴壁情况.结果 TMP能明显抑制HL-60、ECV-304细胞增殖,且能加强As2O3抑制增殖的作用;VEGF mRNA和蛋白表达在TMP与As2O3联合处理后明显减少;TMP与As2O3均能诱导ECV-304细胞早期凋亡,降低细胞贴壁能力,联合作用后效果均明显增强.结论 TMP联合As2O3抑制APL HL-60细胞增殖作用增强,抗血管新生是其治疗APL的可能机制.

Effect of Tetramethylpyrazine Combined with Arsenic Trioxide on the Proliferation of Cell Line HL-60 in Human Promyelocytic Leukemia

Objective To explore the effect and its mechanism of tetramethylpyrazine (TMP) alone or combined with arsenic trioxide (As2O3) on the proliferation of cell line HL-60 in human promyelocytic leukemia (APL). Methods The cell line HL-60 and ECV-304 were adopted and randomized into the blank group, TMP group, As2O3 group and TMP combined with As2O3 group. Four wells were established in each group. Methylthiazolyldiphenyl-tetrazolium bromide (MTT) was used to detect the proliferation of cell line HL-60. The expression of vascular endothelial growth factor (VEGF) mRNA was detected as well. Trypan blue staining method was used to draw the growth curve of cell line ECV-304. The early apoptosis rate of cell line ECV-304 was detected by flow cytometry. The adhesion degree of cell line ECV-304 was observed by inverted microscope. Results TMP could inhibit the proliferation of cell line HL-60 and ECV-304 and strengthen the effect of As2O3. The expression of VEGF mRNA and protein was significantly decreased after the combined treatment. Both TMP and As2O3 could induce the early apoptosis of cell line ECV-304 and reduce the ability of cell adhesion, and the combined treatment could strengthen the effect. Conclusion The combination of TMP and As2O3 has a synergistic effect on inhibiting cell proliferation. It is initially confirmed that anti-angiogenesis is the therapeutic mechanism for APL.