Solov'eva M E, Akatov V S, Leshchenko V V, Kudriavtsev A A
Institute of Theoretical and Experimental Biophysics, Russian Academy of Sciences, Pushchino, Russia.
Izv Akad Nauk Ser Biol. 1998 Mar-Apr(2):194-9.
We studied in vitro myeloma cell death in a serum-free medium. Spectrophotometric assessment of DNA fragmentation, flow cytometry, and staining by Hoechst dye and ethidium bromide indicate that by the beginning of the third day cell death largely followed the apoptotic pathway. We studied dynamics of intracellular pH (pHi) during the cell death and the relationship between the pHi and apoptosis induction during cultivation in media with different pH. We have shown that decreasing pHi in the course of cell death is a consequence rather than the cause of myeloma cell death. Apoptosis took place at cultivation in the media with pH 6.3 and 8.1; the corresponding pHi values were 6.5 and 7.2, respectively. In the presence of Ca-ionophore A23187 we observed appearance of the cells with aberrant distribution of chromatin and fragmented nucleus; however calcium binding in the media with 5-10 mM EDTA induced even more pronounced nuclei fragmentation. This may indicate that both increased and decreased concentration of intracellular calcium induce NS(O) myeloma cell apoptosis.
我们在无血清培养基中研究了体外骨髓瘤细胞死亡情况。通过分光光度法评估DNA片段化、流式细胞术以及用Hoechst染料和溴化乙锭染色表明,到第三天开始时,细胞死亡在很大程度上遵循凋亡途径。我们研究了细胞死亡过程中细胞内pH值(pHi)的动态变化以及在不同pH值培养基中培养期间pHi与凋亡诱导之间的关系。我们已经表明,细胞死亡过程中pHi的降低是骨髓瘤细胞死亡的结果而非原因。在pH值为6.3和8.1的培养基中培养时发生了凋亡;相应的pHi值分别为6.5和7.2。在存在钙离子载体A23187的情况下,我们观察到出现了染色质分布异常和细胞核碎片化的细胞;然而,在含有5 - 10 mM EDTA的培养基中结合钙会诱导更明显的细胞核碎片化。这可能表明细胞内钙浓度的增加和降低均会诱导NS(O)骨髓瘤细胞凋亡。