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目的探讨偏钒酸钠(NaVO3)对HL-60细胞增殖及凋亡的影响,初步判断该影响与ppGalNac-T2的相关性。方法用不同浓度NaVO3作用HL-60细胞后,采用MTT法观察HL-60细胞生长抑制情况,琼脂糖凝胶电泳检测DNA条带,Hoechst 33258染色后荧光显微镜观察细胞核的变化,流式细胞术(FCM)检测细胞凋亡率,分子对接与分子动力学模拟初步判断钒对糖基转移酶ppGalNac-T2活性的影响。结果 MTT检测结果显示,低浓度的Na-VO3促进细胞增殖,高浓度的NaVO3抑制细胞的生长;高浓度NaVO3作用HL-60细胞24 h可见到DNA出现梯形条带,且细胞核呈现核浓缩现象;FCM测定不同浓度NaVO3(5×10-7、1×10-7、1×10-8、1×10-9、1×10-11mol/L)处理HL-60细胞24 h后细胞凋亡率分别是(69.1±2.3)%、(56.3±4.9)%、(39.6±6.5)%、(31.4±1.6)%、(12.2±3.4)%,明显高于HL-60细胞自然凋亡率(均P<0.05);VO3可以与N-乙酰氨基半乳糖转酶2(ppGalNac-T2)的活性中心相关结构形成氢键,并影响ppGalNAc-T2催化相关柔性结构的构象变化,进而影响底物进入和离开酶的活性中心,从而抑制酶的催化功能。结论 NaVO3可以诱导HL-60细胞凋亡,该作用可能与ppGalNAc-T2活性改变相关。
Objective To investigate the effects of sodium metavanadate (NaVO3) on the proliferation and apoptosis of HL-60 cells and to determine the correlation between this effect and ppGalNac-T2. Methods HL-60 cells were treated with different concentrations of NaVO3. MTT assay was used to observe the growth inhibition of HL-60 cells. The DNA bands were detected by agarose gel electrophoresis. Hoechst 33258 staining was used to observe the changes of nuclei. Flow cytometry FCM) to detect the apoptosis rate, molecular docking and molecular dynamics simulation to determine the effect of vanadium on glycosyltransferase ppGalNac-T2 activity. Results MTT results showed that low concentration of Na-VO3 promoted cell proliferation and high concentration of NaVO3 inhibited the growth of cells. High concentration of NaVO3 induced DNA ladder in HL-60 cells for 24 h, and the nuclei showed nuclear condensation. The apoptosis rate of HL-60 cells treated with NaVO3 (5 × 10-7, 1 × 10-7, 1 × 10-8, 1 × 10-9, 1 × 10-11mol / L) (69.1 ± 2.3)%, (56.3 ± 4.9)%, (39.6 ± 6.5)%, (31.4 ± 1.6)% and (12.2 ± 3.4)%, respectively, which were significantly higher than those of HL-60 cells P <0.05). VO3 could form hydrogen bond with the active center structure of N-acetylgalactosaminyl 2 (ppGalNac-T2) and influence the conformational change of ppGalNAc-T2-catalyzed flexible structure, Leave the active center of the enzyme, thereby inhibiting the catalytic function of the enzyme. Conclusion NaVO3 can induce HL-60 cell apoptosis, which may be related to the changes of ppGalNAc-T2 activity.