Yoshihara K, Tsuyuki M, Itaya A, Tanaka Y, Kamiya T
Department of Biochemistry, Nara Medical University, Japan.
Mol Cell Biochem. 1994 Jun 29;135(2):143-51. doi: 10.1007/BF00926517.
HL-60 cells transferred from serum-supplemented to serum-free culture medium initially bound to culture plate tightly and then released from the plate on increasing the culture time and resumed exponential growth after about 8 h lag. At the initial stage of the culture, the cells became extremely sensitive to 3-aminobenzamide, a potent inhibitor of poly (ADP-ribose) polymerase, and, at 1 mM, 80 to 90% of the cells were lysed within 20 h, whereas the inhibitor was totally ineffective on the cell growth in serum-supplemented medium at the concentration. Non-inhibitory analogs of the inhibitor were ineffective. Assay of poly(ADP-ribose) polymerase activity in permeable cells indicated that a transient activation of the enzyme occurred during the culture in serum-free medium (the maximum activation was observed at 8 h of the culture). The cells conditioned in serum-free medium for 24 h acquired significant resistancy to the inhibitor. A low concentration of fibronectin (5 to 10 micrograms/ml) and a relatively high concentration of bovine serum albumin (0.5 to 1 mg/ml) effectively blocked the cell attachment to plate and also the 3-aminobenzamide-induced cell lysis. These results suggest that poly(ADP-ribose) polymerase is involved in a process essential for HL-60 cells to adapt to a serum-deprived growth condition.
从补充血清的培养基转移至无血清培养基的HL-60细胞，最初紧密附着于培养板，随后随着培养时间延长从培养板上脱离，经过约8小时的延迟后恢复指数生长。在培养初期，细胞对聚（ADP-核糖）聚合酶的强效抑制剂3-氨基苯甲酰胺变得极为敏感，在1 mM浓度下，80%至90%的细胞在20小时内裂解，而该抑制剂在该浓度下对补充血清培养基中的细胞生长完全无效。该抑制剂的非抑制性类似物也无效。对可渗透细胞中聚（ADP-核糖）聚合酶活性的测定表明，在无血清培养基培养期间该酶会发生短暂激活（在培养8小时时观察到最大激活）。在无血清培养基中培养24小时的细胞对该抑制剂获得了显著抗性。低浓度的纤连蛋白（5至10微克/毫升）和相对高浓度的牛血清白蛋白（0.5至1毫克/毫升）可有效阻断细胞与培养板的附着以及3-氨基苯甲酰胺诱导的细胞裂解。这些结果表明，聚（ADP-核糖）聚合酶参与了HL-60细胞适应血清剥夺生长条件所必需的一个过程。
