Montesano Gesualdi N, Chirico G, Pirozzi G, Costantino E, Landriscina M, Esposito F
Dipartimento di Biochimica e Biotecnologie Mediche, Università di Napoli Federico II, Naples, Italy.
Stress. 2007 Nov;10(4):342-50. doi: 10.1080/10253890701314863.
TRAP-1 is a mitochondrial heat shock protein (HSP), recently identified in Saos-2 osteosarcoma cells adapted to mild oxidative stress induced by diethylmaleate (DEM). TRAP-1 mRNA expression is increased in DEM-adapted cells as well as in tumor cells resistant to 5-fluorouracil and to platin derivatives. Since a strong decrease of TRAP-1 protein levels, upon cisplatin treatment, is observed only in controls but not in the DEM-adapted counterpart, a possible role for this protein in the development of resistant phenotypes could be hypothesized. To characterize the protective role of TRAP-1 against oxidative stress and apoptosis, stable transfectants were generated and characterized for their response to different stress types. These stable clones expressing constitutively high TRAP-1 levels: (i) are more resistant to H2O2-induced DNA damage and to apoptosis by cisplatin; (ii) contain higher reduced glutathione (GSH) levels than control cells; and (iii) do not release the apoptosis-inducing factor into the nucleus upon cisplatin treatment. Furthermore, high TRAP-1 levels interfere with caspase 3 activation. These results confirm the anti-apoptotic role of TRAP-1, and suggest that increased expression of this mitochondrial HSP in DEM-adapted and chemoresistant cells could be part of a pro-survival signaling pathway aimed to evade toxic effects of oxidants and anticancer drugs.
TRAP-1是一种线粒体热休克蛋白(HSP),最近在适应由马来酸二乙酯(DEM)诱导的轻度氧化应激的Saos-2骨肉瘤细胞中被鉴定出来。在适应DEM的细胞以及对5-氟尿嘧啶和铂衍生物耐药的肿瘤细胞中,TRAP-1 mRNA表达增加。由于仅在对照细胞中观察到顺铂处理后TRAP-1蛋白水平大幅下降,而在适应DEM的对应细胞中未观察到,因此可以推测该蛋白在耐药表型的发展中可能发挥作用。为了表征TRAP-1对氧化应激和细胞凋亡的保护作用,构建了稳定转染子,并对其对不同应激类型的反应进行了表征。这些持续高表达TRAP-1水平的稳定克隆:(i)对H2O2诱导的DNA损伤和顺铂诱导的细胞凋亡更具抗性;(ii)比对照细胞含有更高水平的还原型谷胱甘肽(GSH);(iii)在顺铂处理后不会将凋亡诱导因子释放到细胞核中。此外,高TRAP-1水平会干扰半胱天冬酶3的激活。这些结果证实了TRAP-1的抗凋亡作用,并表明在适应DEM和具有化疗抗性的细胞中这种线粒体HSP表达增加可能是旨在逃避氧化剂和抗癌药物毒性作用的促生存信号通路的一部分。