Primary cell cultures arising from normal kidney and renal cell carcinoma retain the proteomic profile of corresponding tissues
Primary cell cultures arising from normal kidney and renal cell carcinoma retain the proteomic profile of corresponding tissues
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DOI：
10.1021/pr050002o
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发表时间：
2005-09-01
影响因子：
4.4
通讯作者：
Mocarelli, P
中科院分区：
文献类型：
--
作者：
Perego, RA;Bianchi, C;Mocarelli, P
肾细胞癌(RCC)组织是由肿瘤和正常细胞组成的混合体，这使得蛋白质组分析变得复杂。我们研究的目的是探讨建立肾细胞癌和肾皮质的原代细胞培养是否可行，以保持组织表型以及更均匀和丰富的细胞学材料。建立了17例手术患者的14种原代培养物(82.3%)，并对其进行了形态、生长速度、免疫细胞化学和分子分析，包括实时定量聚合酶链式反应、Western blotting、双向电泳法和质谱仪。培养显示90%的细胞角蛋白阳性的上皮细胞。在原代肿瘤培养中，锰超氧化物歧化酶和热休克蛋白27的分子表型与肿瘤组织中发现的相同，但表达增加，亚型增加。此外，从皮质或肾细胞癌培养物的2-DE凝胶上切下的斑点中，有27种特异蛋白质及其异构体与2-DE组织皮质参考图上鉴定的蛋白质及其异构体相对应，这表明这些原代培养物保持了相应组织的蛋白质组学特征。
Renal cell carcinoma (RCC) tissue is composed of a mixture of neoplastic and normal cells, which complicate proteome analysis. The aim of our study was to investigate whether it is feasible to establish primary cell cultures of RCC and of renal cortex maintaining the tissue phenotype along with a more homogeneous and enriched cytological material. Fourteen (82.3%) primary cultures from 17 surgical cases were established and characterized by morphology, growth rate, immunocytochemistry, and molecular analysis performed by Real-time PCR, Western blotting, two-dimensional electrophoresis (2-DE), and mass spectrometry. Cultures showed >90% cytokeratine-positive epithelial cells. In primary tumor cultures, the molecular phenotype of manganese superoxide dismutase and heat shock protein 27 was the same as that found in tumor tissues with overexpression and increased number of isoforms. Moreover, 27 out 28 specific proteins and their isoforms, present in spots excised from 2-DE gel of cortex or RCC cultures, corresponded to those identified on the 2-DE tissue cortex reference map, suggesting that these primary cultures retain the proteomic profile of the corresponding tissues.