Department of Developmental Biology, Institute of Biochemistry, Mokslininku st. 12, 08662, Vilnius, Lithuania.
In Vitro Cell Dev Biol Anim. 2010 Jun;46(6):547-59. doi: 10.1007/s11626-010-9296-0. Epub 2010 Mar 12.
The modifications of intracellular redox balance leads to important cellular changes in many cell types. Here, a causal relationship among redox state, granulocytic differentiation induced by all-trans retinoic acid (RA) or dibutyryl cAMP (dbcAMP) and apoptosis have been studied in the human acute promyelocytic leukaemia HL-60 cells. The modulation of intracellular reactive oxygen species levels by D: , L: -buthionine-(S, R) sulfoximide (BSO), and N: -acetyl-L: -cysteine (NAC) caused inducer- and time-dependent or stage-specific effects on HL-60 cell growth inhibition, differentiation and subsequent apoptosis. The presence of BSO during the commitment stage suppressed RA-but not dbcAMP-mediated differentiation, while NAC inhibited both. BSO alone and in combination with RA or dbcAMP-induced apoptosis, which was prevented by NAC in dbcAMP-but not in RA-treated cells. Using protein kinase C inhibitor, calphostin C, cross-talk effects between the intracellular redox state and PKC signalling was identified by demonstrating inducer-dependent changes in cell differentiation or apoptosis, which were associated with the changes in DNA-NF-kappaB binding activity. These observations suggest a critical role of redox state in determining HL-60 cell behaviour and provide new insights into the complex effects of redox perturbations on the intracellular signalling network via the involvement of PKC and NF-kappaB.
细胞内氧化还原平衡的改变会导致许多细胞类型发生重要的细胞变化。在这里，我们研究了全反式视黄酸（RA）或二丁酰环腺苷酸（dbcAMP）诱导的粒细胞分化与细胞凋亡之间的因果关系在人急性早幼粒细胞白血病 HL-60 细胞中的关系。通过 D:, L: -丁硫氨酸-(S, R)亚砜（BSO）和 N: -乙酰基-L: -半胱氨酸（NAC）调节细胞内活性氧水平，对 HL-60 细胞生长抑制、分化和随后的凋亡产生诱导剂和时间依赖性或阶段特异性影响。在承诺阶段存在 BSO 会抑制 RA 但不抑制 dbcAMP 介导的分化，而 NAC 则抑制两者。BSO 单独以及与 RA 或 dbcAMP 联合诱导细胞凋亡，NAC 可防止 dbcAMP 处理的细胞发生凋亡，但不能防止 RA 处理的细胞发生凋亡。使用蛋白激酶 C 抑制剂 calphostin C，通过证明细胞分化或凋亡的诱导剂依赖性变化，确定了细胞内氧化还原状态与蛋白激酶 C 信号之间的交叉对话效应，这些变化与 DNA-NF-κB 结合活性的变化有关。这些观察结果表明，氧化还原状态在决定 HL-60 细胞行为方面起着关键作用，并为通过蛋白激酶 C 和 NF-κB 的参与，氧化还原扰动对细胞内信号网络的复杂影响提供了新的见解。
