Glutathione induces cellular resistance against cationic dinuclear platinum anticancer drugs

摘要

The sulfur-containing tripeptide glutathione (GSH) is one of the most abundant molecules in cells. Elevated levels of GSH render some types of cancer cells resistant against well-known platinum anti-cancer drugs such as cisplatin and carboplatin. Platinum complexes are often very reactive towards the cysteine residue of GSH, which detoxifies these compounds by a rapid binding mechanism. Clearly, this resistance mechanism poses a severe obstacle to any new platinum drugs designed to overcome cisplatin resistance. In the present study the cytotoxicity of dinuclear platinum compounds of the 1, 1 /t,t type, as developed by Farrell, is determined in human ovarium A2780 cells and in the cisplatin-resistant cell line A2780cisR, which possesses elevated levels of GSH. Further, the effect of depletion of GSH levels by L-buthionine-S,R-sulfoxiniine (L-BSO) in A2780cisR was investigated. The experiments show that detoxification by GSH is an effective resistance mechanism against dinuclear platinum compounds. However, the dinuclear complexes are less sensitive towards detoxification compared to cisplatin. This is probably because of the rapid binding of dinuclear cationic complexes to DNA. Compared to cisplatin, the rapid binding to DNA reduces the time during which the drug molecules are exposed to GSH in the cytosol. The reaction of a representative dinuclear compound with glutathione (pH 7, 37 degreesC) was studied in detail by Pt-195 NMR. The dinuclear complex BBR3005 ([trans-PtCl2(NH3)(2)(mu-H2N(CH2)(6)NH2)](2+), abbreviated as 1,1/t,t n=6), follows different pathways in the reaction with GSH, depending on the molar ratio of the reactants. When reacted in stoichometric amounts (1:1), first a chloride on each platinum is replaced by a sulfur, forming a PtN3S product at -2977 ppm. After 2-3 h, this intermediate reacts further to form a sulfur-bridged N3Pt-S-PtN3 species as the main product at -2811 ppm. When BBR3005 is reacted with GSH in a ratio of 1:4, the sulfur-bridged species is not observed. Instead, the final product is trans-Pt(GS)(2)(NH3)(2) (at -3215 ppm); the same product appears if GSH is reacted with trans-PtCl2(NH3)(2). Apparently, GSH first replaces the chlorides and subsequently degrades the dinuclear compound by replacement of the diaminealkyl linker.