Unwin Richard D, Craven Rachel A, Harnden Patricia, Hanrahan Sarah, Totty Nick, Knowles Margaret, Eardley Ian, Selby Peter J, Banks Rosamonde E
Cancer Research UK Clinical Unit, St. James's University Hospital, Beckett Street, Leeds LS9 7TF, UK.
Proteomics. 2003 Aug;3(8):1620-32. doi: 10.1002/pmic.200300464.
Renal cell carcinoma (RCC) is the tenth most common cancer although the incidence is increasing. The main clinical problems stem from the relatively late presentation of many patients due to the often asymptomatic nature of the illness, and the relative insensitivity of metastatic disease to conventional chemotherapy and radiotherapy. Despite increasing knowledge of some of the genetic changes underlying sporadic renal cancer such as those involving the Von Hippel Lindau (VHL) gene, many of the underlying pathophysiological changes are ill-defined and there remains a need for the identification of disease markers for use in diagnosis and prognosis or as potential therapeutic targets. This study has used a proteomic approach, based on two-dimensional gel electrophoresis and mass spectrometry, to compare the protein profiles of conventional RCC tissue with patient-matched normal kidney cortex. Sequencing of 32 protein spots with significantly increased expression in RCC samples (>/= 4/6 patients) and 41 proteins whose levels decreased (6/6 patients) confirmed several previously known RCC-associated changes such as increases in Mn-superoxide dismutase, lactate dehydrogenase-A, aldolase A and C, pyruvate kinase M2, and thymidine phosphorylase. Additionally, several previously unknown changes were identified, including increased expression of three members of the annexin family and increased levels of the actin depolymerisation factor cofilin. The Warburg effect was also demonstrated with the identification of increases in proteins involved in the majority of steps in the glycolytic pathway and decreases in the gluconeogenic reactions, together with a parallel decrease in several mitochondrial enzymes. A number of the alterations seen were further confirmed in additional samples by immunohistochemistry, Western blotting, and laser capture microdissection.
肾细胞癌（RCC）是第十大常见癌症，尽管其发病率正在上升。主要临床问题源于许多患者就诊时相对较晚，这是由于该疾病通常无症状，以及转移性疾病对传统化疗和放疗相对不敏感。尽管对散发性肾癌的一些潜在基因变化（如涉及冯·希佩尔-林道（VHL）基因的变化）的了解不断增加，但许多潜在的病理生理变化仍不明确，仍需要鉴定用于诊断和预后或作为潜在治疗靶点的疾病标志物。本研究采用了一种基于二维凝胶电泳和质谱的蛋白质组学方法，比较了传统RCC组织与患者匹配的正常肾皮质的蛋白质谱。对RCC样本中表达显著增加（≥4/6患者）的32个蛋白点和水平降低的41个蛋白（6/6患者）进行测序，证实了一些先前已知的与RCC相关的变化，如锰超氧化物歧化酶、乳酸脱氢酶-A、醛缩酶A和C、丙酮酸激酶M2和胸苷磷酸化酶的增加。此外，还发现了一些先前未知的变化，包括膜联蛋白家族三个成员的表达增加和肌动蛋白解聚因子cofilin水平的增加。通过鉴定糖酵解途径中大多数步骤相关蛋白的增加和糖异生反应的减少，以及几种线粒体酶的平行减少，也证明了瓦伯格效应。通过免疫组织化学、蛋白质印迹和激光捕获显微切割在其他样本中进一步证实了所观察到的一些变化。
