Apoptotic and non-apoptotic cell death induced by cis and trans analogues of a novel ammine(cyclohexylamine)dihydroxodichloroplatinum(IV) complex.
Apoptotic and non-apoptotic cell death induced by cis and trans analogues of a novel ammine(cyclohexylamine)dihydroxodichloroplatinum(IV) complex.
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DOI：
10.1038/bjc.1996.486
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发表时间：
1996-10
影响因子：
8.8
通讯作者：
Kelland, L R
中科院分区：
文献类型：
--
作者：
O'Neill, C F;Ormerod, M G;Robertson, D;Titley, J C;Cumber-Walsweer, Y;Kelland, L R
以前已经证明顺铂诱导CH 1人卵巢癌细胞系的凋亡。这项研究表明，两种新的铂（Pt）类似物JM 149和JM 335，这是顺式和反式几何分别氨（环己胺）dihydroxodichloroPt（IV），启动细胞凋亡在该细胞系在生理相关的浓度（IC 50值2小时药物暴露JM 149和JM 335分别为35.3 μ M和18.7 μ M）。虽然在等摩尔药物浓度下，暴露于JM 335与JM 149后总铂-DNA加合物的水平高2倍，但在等毒性浓度下，水平相似（分别为80 pmol Pt mg-1 DNA与70 pmol Pt mg-1 DNA）。在与2 × IC 50的两种药物孵育2小时后，细胞以时间依赖性方式变圆并分离，但细胞凋亡的动力学对于JM 335更快。大多数分离的细胞表现出与凋亡相关的形态，这进一步得到了从这些细胞制备的DNA裂解物中检测到的50 kb片段的存在的支持。JM 149在一系列浓度（2 x、5 x和10 x IC 50）下诱导细胞凋亡，在所有浓度下均检测到50 kb DNA片段。然而，与此形成鲜明对比的是，JM 335在10 x IC 50下未能引起细胞凋亡，脱离的细胞既没有显示出细胞凋亡形态，也没有可检测到的50 kb DNA片段。此外，这些分离的细胞显示出广泛的囊泡形成的证据，而DNA在外观上保持正常，因此似乎已经通过非凋亡模式死亡。与在2 x IC 50下观察到的片段相比，在5 x IC 50的JM 335下诱导的细胞凋亡程度似乎也较低，这通过较低强度的50 kb片段证明。这些药物的主要细胞周期效应（2 × IC 50）是S期穿越的减缓，在此期间似乎发生了大部分但不是全部的细胞凋亡。然而，在5 x IC 50时，JM 335细胞在细胞周期的所有阶段都出现冷冻，从G1到S几乎没有进展，伴随着G2期细胞的积聚，表明G2/M阻滞。细胞周期效应的这种差异可能是该浓度下细胞凋亡水平降低以及较高浓度下细胞凋亡失败的原因。这些数据表明，铂类药物的性质（以及因此产生的DNA损伤的性质）可能对确定该细胞系中细胞死亡的速率和机制具有重要意义。用反式复合物JM 335观察到的细胞死亡效应可能与该细胞系中DNA单链断裂的诱导相关。
It has been previously demonstrated that cisplatin induces apoptosis in the CH1 human ovarian carcinoma cell line. This study demonstrates that two novel platinum (Pt) analogues JM149 and JM335, which are the cis and trans geometry respectively of ammine(cyclohexylamine)dihydroxodichloroPt(IV), initiate apoptosis in this cell line at physiologically relevant concentrations (IC50 values 2 h drug exposure were 35.3 microM for JM149 and 18.7 microM for JM335). While at equimolar drug concentrations there was a 2-fold higher level of total platinum-DNA adducts following exposure to JM335 vs JM149, at equitoxic concentrations, levels were similar (80 vs 70 pmol Pt mg-1 DNA respectively). Following a 2 h incubation with 2 x IC50 of both drugs, cells rounded up and detached in a time-dependent manner but with the kinetics of apoptosis being more rapid for JM335. The majority of detached cells exhibited morphology associated with apoptosis which was further supported by the presence of a 50 kb fragment detected in DNA lysates prepared from these cells. JM149 induced apoptosis across a range of concentrations (2 x, 5 x and 10 x IC50) with a 50 kb DNA fragment being detected at all concentrations. However, in marked contrast to this, JM335 failed to cause apoptosis at 10 x IC50, the detached cells neither displaying apoptotic morphology nor a detectable 50 kb DNA fragment. Moreover, these detached cells showed evidence of extensive vesiculation while the DNA remained normal in appearance and thus appeared to have died by a non-apoptotic mode. Apoptosis also appeared to be induced to a lesser extent at 5 x IC50 of JM335 as demonstrated by a less intense 50 kb fragment compared with that seen at 2 x IC50. The main cell cycle effect of these drugs (at 2 x IC50) was a slowdown in S-phase traverse during which most but not all of the apoptosis appeared to occur. However, at 5 x IC50 of JM335 cells appeared frozen in all phases of the cell cycle with little progress from G1 to S accompanied by a build-up of cells in G2 indicative of a G2/M block. This difference in cell cycle effect may account for the reduced level of apoptosis at this concentration and a failure to engage apoptosis at higher concentrations. These data suggest that the nature of the platinum drug (and consequently, the nature of resultant DNA damage) may have important implications in determining the rate and mechanism of cell death in this cell line. The cell death effects observed with the trans complex JM335 may correlate with the induction of DNA single-strand breaks in this cell line.