Disparate cytogenetic responses of peripheral blood and spleen lymphocytes to ethenoadenine nucleotides in vitro; maximal expression in splenic lymphocytes under conditions of enhanced membrane permeabilization.
Disparate cytogenetic responses of peripheral blood and spleen lymphocytes to ethenoadenine nucleotides in vitro; maximal expression in splenic lymphocytes under conditions of enhanced membrane permeabilization.
复制标题
体外外周血和脾淋巴细胞对乙烯腺嘌呤核苷酸的不同细胞遗传学反应；
DOI：
10.1093/carcin/11.4.571
复制
发表时间：
1990
期刊：
影响因子：
4.7
通讯作者：
Conner,MK
中科院分区：
文献类型：
--
作者：
Modzelewski,RA;Conner,MK
外源性1，N6-亚乙基腺苷三磷酸（ε-ATP）和1，N6-亚乙基脱氧腺苷三磷酸（ε-dATP）是小鼠脾淋巴细胞姐妹染色单体交换（SCE）的强诱导剂，但对体外培养的外周血淋巴细胞无诱导作用。数据表明，脾淋巴细胞膜对ε-ATP和εdATP的瞬时摄取具有比血液淋巴细胞更高的固有渗透性。本文研究了培养基pH和二价阳离子对ε-ATP脉冲处理的脾细胞SCE频率和染色体畸变的影响。在pH 8.0的无Ca ~（2+）/Mg ~（2+）的Hank平衡盐溶液（HBSS）中观察到最显著的响应。在后一种条件下，重复小鼠淋巴细胞的SCE和染色体畸变反应（平均值± SD）分别为69.4 ± 13.1 SCE/细胞和49 ± 8.5%畸变细胞。染色体畸变包括多种复杂的断裂和重排。在HBSS含有钙（0.575 mM）和镁（0.4 mM）的浓度相当于那些在RPMI 1640，最大SCE和畸变反应的31.8和28%，观察到在pH 6.0处理的细胞。同样，在pH 6.0的RPMI培养基中处理的脾细胞中，SCE频率最高（46 ± 1.6 SCE/cen），畸变细胞百分比最高（8 ± 1.4%）。SCE和畸变随pH值的增加而降低，在任一媒体含有二价阳离子。在无Ca 2 +/Mg 2+的HBSS中，在pH 6.0下观察到最高的有丝分裂指数和最快的细胞周期。有丝分裂指数显着下降，在pH 7.4，但恢复相当大，在pH 8.0，尽管含有异常染色体的细胞的频率很高。最显著的细胞毒性发生在pH 6.0的HBSS含有Ca ~（2+）和Mg ~（2+）。在较高pH和RPMI培养基中，细胞毒性明显降低。对照细胞在无Ca ~（2+）/Mg ~（2+）的HBSS培养基中，在高pH值（8.0）下脉冲处理后，获得了最佳生长条件。由于外源性ε-ATP具有显著的细胞遗传学毒性，以及细胞内ATP的普遍存在和生物学重要性，后者应被视为致癌物质亲电代谢产物形成加合物的潜在靶点。
Exogenously supplied l,N6-ethenoadenosine triphosphate (ε-ATP) and 1,N6-ethenodeoxyadewsine triphosphate (ε-dATP) are potent inducers of sister chromatid exchanges (SCEs) in murine spleen lymphocytes but not in peripheral blood lymphocytes culturedin vitro. Data suggest that spleen lymphocyte membranes are inherently more permeable than blood lymphocytes to transient uptake of ε-ATP and εdATP. The effect of media pH and divalent cations on SCE frequency and chromosomal aberrations in spleen cells pulse-treated with ε-ATP were studied. The most dramatic responses were observed at pH 8.0 in ca2+/Mg2+-free Hank's balanced salt solution (HBSS). Under the latter conditions, SCE and chromosomal aberration responses (mean ± SD) of lymphocytes from replicate mice were 69.4 ± 13.1 SCE/cell and 49 ± 8.5% of cells with aberrations respectively. Chromosomal aberrations included multiple complex breakage and rearrangements. In HBSS containing ca2+(0.575 mM) and Mg2+(0.4 mM) in concentrations equivalent to those in RPMI 1640, maximum SCE and aberration responses of 31.8 and 28% were observed in cells treated at pH 6.0. Similarly, maximum SCE frequencies (46 ± 1.6 SCE/cen) and percentage of cells with aberrations (8 ± 1.4%) were present in spleen cells treated at pH 6.0 in RPMI media. SCEs and aberrations decreased with increasing pH in either media containing divalent cations. In Ca2+/Mg2+-free HBSS, the highest mitotic index and fastest cell cycling were seen at pH 6.0. Mitotic indices dropped dramatically at pH 7.4 but recovered considerably at pH 8.0, in spite of a high frequency of cells containing aberrant chromosomes. The most dramatic cytotoxicity occurred at pH 6.0 in HBSS containing ca2+and Mg2+. Decreased cytotoxicity was apparent at higher pH and in RPMI medium. Conditions for optimal growth of control cells were obtained following pulsetreatment in Ca2+/Mg2+-free HBSS medium at high pH (8.0). Because of the dramatic cytogenetic toxicity of exogenously supplied ε-ATP, and the ubiquitous occurrence and biological importance of intracellular ATP, the latter should be considered a potential target for adduct formation by electrophilic metabolites of carcinogenic agents.