Objective: To investigate the effects of methionine restriction on proliferation, cell cycle and apoptosis of human acute leukemia cells.
Methods: Cell Counting Kit-8 (CCK-8) assay was used to detect the effect of methionine restriction on HL-60 and Jurkat cells proliferation. The effect of methionine restriction on cell cycle of HL-60 and Jurkat cells was examined by PI staining. Annexin V-FITC / PI double staining was applied to detect apoptosis of HL-60 and Jurkat cells following methionine restriction. The expression of cell cycle-related proteins cyclin B1, CDC2 and apoptosis-related protein Bcl-2 was evaluated by Western blot assay.
Results: Methionine restriction significantly inhibited the proliferation of HL-60 and Jurkat cells in a time-dependent manner (HL-60: r =0.7773, Jurkat: r =0.8725), arrested the cells at G2/M phase (P < 0.001), and significantly induced apoptosis of HL-60 and Jurkat cells (HL-60: P < 0.001; Jurkat: P < 0.05). Furthermore, Western blot analysis demonstrated that methionine restriction significantly reduced the proteins expression of Cyclin B1 (P < 0.05), CDC2 (P < 0.01) and Bcl-2 (P < 0.001) in HL-60 and Jurkat cells.
Conclusion: Acute leukemia cells HL-60 and Jurkat exhibit methionine dependence. Methionine restriction can significantly inhibit the proliferation, promote cell cycle arrest and induce apoptosis of HL-60 and Jurkat cells, which suggests that methionine restriction may be a potential therapeutic strategy for acute leukemia.
题目: 甲硫氨酸限制对人急性白血病细胞增殖、周期及凋亡的影响.
目的: 探讨甲硫氨酸限制对人急性白血病细胞增殖、周期和凋亡的影响。.
方法: 采用Cell Counting Kit -8（CCK-8）法检测甲硫氨酸限制对急性白血病细胞HL-60和Jurkat增殖的影响；PI单染法检测甲硫氨酸限制对HL-60和Jurkat细胞周期的影响；Annexin V-FITC/PI双染法检测甲硫氨酸限制后HL-60和Jurkat细胞凋亡的情况；Western blot检测甲硫氨酸限制后细胞周期相关蛋白Cyclin B1和CDC2以及凋亡相关蛋白Bcl-2的表达。.
结果: 甲硫氨酸限制对HL-60和Jurkat细胞增殖具有显著抑制作用且呈时间依赖性（HL-60: r =0.7773；Jurkat: r =0.8725），并将急性白血病细胞HL-60和Jurkat阻滞在G2/M期（P <0.001），能够显著诱导HL-60和Jurkat细胞凋亡（HL-60: P <0.001；Jurkat: P <0.05）。Western blot结果表明，甲硫氨酸限制显著降低HL-60和Jurkat细胞内周期相关蛋白Cyclin B1（P <0.05）和CDC2（P <0.01）及凋亡相关蛋白Bcl-2（P <0.001)的表达。.
结论: 急性白血病细胞HL-60和Jurkat具有甲硫氨酸依赖性，甲硫氨酸限制能够显著抑制HL-60和Jurkat细胞的增殖，促进HL-60和Jurkat细胞发生细胞周期阻滞和凋亡，提示甲硫氨酸限制可能是治疗急性白血病的一种潜在治疗策略。.
Keywords: acute leukemia; apoptosis; cell cycle; homocysteine; methionine; proliferation.