Yang Youfeng, Valera Vladimir A, Padilla-Nash Hesed M, Sourbier Carole, Vocke Cathy D, Vira Manish A, Abu-Asab Mones S, Bratslavsky Gennady, Tsokos Maria, Merino Maria J, Pinto Peter A, Srinivasan Ramaprasad, Ried Thomas, Neckers Len, Linehan W Marston
Urologic Oncology Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, 10 Center Dr., MSC 1107, Bldg 10 CRC, Room 1-5942, Bethesda, MD 20892-1107.
Cancer Genet Cytogenet. 2010 Jan 1;196(1):45-55. doi: 10.1016/j.cancergencyto.2009.08.018.
Energy deregulation and abnormalities of tumor cell metabolism are critical issues in understanding cancer. Hereditary leiomyomatosis renal cell carcinoma (HLRCC) is an aggressive form of RCC characterized by germline mutation of the Krebs cycle enzyme fumarate hydratase (FH), and one known to be highly metastatic and unusually lethal. There is considerable utility in establishing preclinical cell and xenograft models for study of disorders of energy metabolism, as well as in development of new therapeutic approaches targeting of tricarboxylic acid (TCA) cycle enzyme-deficient human cancers. Here we describe a new immortalized cell line, UOK 262, derived from a patient having aggressive HLRCC-associated recurring kidney cancer. We investigated gene expression, chromosome profiles, efflux bioenergetic analysis, mitochondrial ultrastructure, FH catabolic activity, invasiveness, and optimal glucose requirements for in vitro growth. UOK 262 cells have an isochromosome 1q recurring chromosome abnormality, i(1)(q10), and exhibit compromised oxidative phosphorylation and in vitro dependence on anaerobic glycolysis consistent with the clinical manifestation of HLRCC. The cells also display glucose-dependent growth, an elevated rate of lactate efflux, and overexpression of the glucose transporter GLUT1 and of lactate dehydrogenase A (LDHA). Mutant FH protein was present primarily in edematous mitochondria, but with catalytic activity nearly undetectable. UOK 262 xenografts retain the characteristics of HLRCC histopathology. Our findings indicate that the severe compromise of oxidative phosphorylation and rapid glycolytic flux in UOK 262 are an essential feature of this TCA cycle enzyme-deficient form of kidney cancer. This tumor model is the embodiment of the Warburg effect. UOK 262 provides a unique in vitro and in vivo preclinical model for studying the bioenergetics of the Warburg effect in human cancer.
能量去调控和肿瘤细胞代谢异常是理解癌症的关键问题。遗传性平滑肌瘤病肾细胞癌(HLRCC)是肾细胞癌的一种侵袭性形式,其特征是三羧酸循环酶富马酸水合酶(FH)的种系突变,并且已知具有高度转移性和异常致死性。建立临床前细胞和异种移植模型用于研究能量代谢紊乱以及开发针对三羧酸(TCA)循环酶缺陷型人类癌症的新治疗方法具有重要意义。在此,我们描述了一种新的永生化细胞系UOK 262,它源自一名患有侵袭性HLRCC相关复发性肾癌的患者。我们研究了基因表达、染色体图谱、外排生物能量分析、线粒体超微结构、FH分解代谢活性、侵袭性以及体外生长的最佳葡萄糖需求。UOK 262细胞具有1号等臂染色体重复染色体异常i(1)(q10),并且表现出氧化磷酸化受损以及体外对无氧糖酵解的依赖性,这与HLRCC的临床表现一致。这些细胞还表现出葡萄糖依赖性生长、乳酸外排率升高以及葡萄糖转运蛋白GLUT1和乳酸脱氢酶A(LDHA)的过表达。突变的FH蛋白主要存在于水肿的线粒体中,但催化活性几乎检测不到。UOK 262异种移植保留了HLRCC组织病理学特征。我们的研究结果表明,UOK 262中氧化磷酸化的严重受损和快速糖酵解通量是这种TCA循环酶缺陷型肾癌的一个基本特征。这个肿瘤模型是瓦伯格效应的体现。UOK 262为研究人类癌症中瓦伯格效应的生物能量学提供了一个独特的体外和体内临床前模型。