Rephaeli A, Aviram A, Rabizadeh E, Shaklai M
Hematology Division, Beilinson Medical Center, Petach Tikva, Israel.
J Cell Physiol. 1990 Apr;143(1):154-9. doi: 10.1002/jcp.1041430121.
Proliferation-associated changes in calcium metabolism were investigated employing the promyelocytic HL-60 and monoblastic U-937 cell lines. The cells were stimulated to proliferate employing mitogenic factors as follows. 1) Transferrin or insulin: HL-60 cells were adjusted for growth in serum-free medium, and 24 h prior to the experiment, the cells were deprived of transferrin or insulin. The re-addition of either one of them stimulated cell proliferation as was evident by increased [3H]-tymidine incorporation activity. Cell proliferation was associated with an enhanced Ca2+ influx rate, measured by 45Ca2+ uptake activity. 2) Granulocyte-monocyte colony-stimulating factor (GM-CSF): addition of GM-CSF to proliferating or quiescent HL-60 cells resulted in increased cell proliferation, which was also accompanied by increased rate of Ca2+ influx. 3) Serum: HL-60 and U-937 were grown for 24 h in serum-depleted medium. Re-addition of serum to the cells was not associated with immediate or delayed change in calcium influx rate but rather with an immediate increase in the cytosolic free calcium concentration, measured employing the fluorescent probe, fura-2AM. This increase was independent of extracellular calcium, unaffected by verapamil, diltiazem, and lanthanum, and associated with enhanced 45Ca2+ efflux. Thus, in all three cases evoked cell proliferation was accompanied by quantitative changes in Ca2+ metabolism. While the transferrin-, insulin-, and GM-CSF-stimulated cell proliferation was accompanied by delayed increases in 45Ca2+ influx, the serum-stimulated cell proliferation was accompanied by an immediate elevation of free cytosolic Ca2+.
利用早幼粒细胞HL - 60和单核细胞U - 937细胞系研究了与增殖相关的钙代谢变化。使用有丝分裂原因子刺激细胞增殖，具体如下：1）转铁蛋白或胰岛素：将HL - 60细胞调整为在无血清培养基中生长，在实验前24小时，去除细胞中的转铁蛋白或胰岛素。重新添加其中任何一种都会刺激细胞增殖，这通过增加的[³H] - 胸腺嘧啶核苷掺入活性得以体现。细胞增殖与通过⁴⁵Ca²⁺摄取活性测量的增强的Ca²⁺内流速率相关。2）粒细胞 - 单核细胞集落刺激因子（GM - CSF）：向增殖或静止的HL - 60细胞中添加GM - CSF会导致细胞增殖增加，这也伴随着Ca²⁺内流速率的增加。3）血清：将HL - 60和U - 937细胞在无血清培养基中培养24小时。向细胞中重新添加血清与钙内流速率的即时或延迟变化无关，而是与使用荧光探针fura - 2AM测量的胞质游离钙浓度的即时增加有关。这种增加与细胞外钙无关，不受维拉帕米、地尔硫䓬和镧的影响，并且与增强的⁴⁵Ca²⁺外流相关。因此，在所有三种引起细胞增殖的情况下，都伴随着Ca²⁺代谢的定量变化。虽然转铁蛋白、胰岛素和GM - CSF刺激的细胞增殖伴随着⁴⁵Ca²⁺内流的延迟增加，但血清刺激的细胞增殖伴随着胞质游离钙的即时升高。
