Pluim Dick, van Waardenburg Robert C A M, Beijnen Jos H, Schellens Jan H M
Division of Experimental Therapy and Medical Oncology, The Netherlands Cancer Institute, Antoni van Leeuwenhoek Hospital, Plesmanlaan 121, 1066CX Amsterdam, The Netherlands.
Cancer Chemother Pharmacol. 2004 Jul;54(1):71-8. doi: 10.1007/s00280-004-0773-6. Epub 2004 Mar 19.
The cytotoxicity, intracellular accumulation and DNA adduct formation of the ruthenium complex imidazolium trans-imidazoledimethylsulfoxide tetrachlororuthenate (ImH[ trans-RuCl(4)(DMSO)Im], Nami-A) were compared in vitro with those of cisplatin in four human tumor cell lines: Igrov-1, 2008, MCF-7, and T47D.
Cytotoxicity was assessed in vitro using a growth inhibition assay. Accumulation was determined by flameless atomic absorption spectroscopy (AAS). GG and AG intrastrand adducts were measured using the (32)P-postlabeling assay.
Nami-A was on average 1053 times less cytotoxic than cisplatin. The cytotoxicity of cisplatin was linearly related to both intracellular platinum accumulation and DNA binding, while the cytotoxicity of Nami-A was significantly related only to DNA binding and not to intracellular ruthenium accumulation. The levels of accumulation of Nami-A measured as ruthenium and of cisplatin measured as platinum were correlated linearly with the incubation concentration over a concentration range of 0 to 600 micro M of both drugs. Ruthenium intracellular accumulation and DNA binding were on average 4.8 and 42 times less, respectively, than those of cisplatin. In addition, the numbers of GG and AG intrastrand adducts induced by Nami-A were 418 and 51 times fewer, respectively. Nami-A and cisplatin had the same binding capacity to calf thymus DNA. Nami-A was 25-40% less bound to cellular proteins than cisplatin.
There was no saturation of the uptake and DNA binding capacity of either Nami-A or cisplatin. Furthermore, the low binding of Nami-A to cellular DNA cannot simply be explained by a lower capacity to bind to DNA, because the absolute level of binding in vitro to calf thymus DNA was the same for Nami-A and cisplatin. Finally, the lower cytotoxicity of Nami-A on a molar basis than that of cisplatin can at least partly be explained by its reduced reactivity to DNA in intact cells.
在四种人类肿瘤细胞系（Igrov-1、2008、MCF-7和T47D）中，对钌配合物咪唑鎓反式-咪唑二甲基亚砜四氯钌酸盐（ImH[trans-RuCl(4)(DMSO)Im]，Nami-A）的细胞毒性、细胞内蓄积及DNA加合物形成与顺铂进行体外比较。
采用生长抑制试验体外评估细胞毒性。通过无火焰原子吸收光谱法（AAS）测定蓄积情况。使用³²P后标记试验测量GG和AG链内加合物。
Nami-A的细胞毒性平均比顺铂低1053倍。顺铂的细胞毒性与细胞内铂蓄积及DNA结合均呈线性相关，而Nami-A的细胞毒性仅与DNA结合显著相关，与细胞内钌蓄积无关。在0至600 μM的两种药物浓度范围内，以钌测量的Nami-A蓄积水平和以铂测量的顺铂蓄积水平与孵育浓度呈线性相关。钌的细胞内蓄积和DNA结合平均分别比顺铂少4.8倍和42倍。此外，Nami-A诱导的GG和AG链内加合物数量分别少418倍和51倍。Nami-A和顺铂对小牛胸腺DNA具有相同结合能力。Nami-A与细胞蛋白的结合比顺铂少25 - 40%。
Nami-A或顺铂的摄取及DNA结合能力均未饱和。此外，Nami-A与细胞DNA的低结合不能简单地用其较低的DNA结合能力来解释，因为Nami-A和顺铂在体外对小牛胸腺DNA结合的绝对水平相同。最后，Nami-A在摩尔基础上比顺铂细胞毒性低至少部分可归因于其在完整细胞中对DNA反应性降低。
