Myung Jae-Kyung, Afjehi-Sadat Leila, Felizardo-Cabatic Maureen, Slavc Irene, Lubec Gert
Department of Pediatrics, Medical University of Vienna, Vienna, Austria.
Proteome Sci. 2004 Dec 14;2(1):8. doi: 10.1186/1477-5956-2-8.
Chaperones (CH) play an important role in tumor biology but no systematic work on expressional patterns has been reported so far. The aim of the study was therefore to present an analytical method for the concomitant determination of several CH in human tumor cell lines, to generate expressional patterns in the individual cell lines and to search for tumor and non-tumor cell line specific CH expression.Human tumor cell lines of neuroblastoma, colorectal and adenocarcinoma of the ovary, osteosarcoma, rhabdomyosarcoma, malignant melanoma, lung, cervical and breast cancer, promyelocytic leukaemia were homogenised, proteins were separated on two-dimensional gel electrophoresis with in-gel digestion of proteins and MALDI-TOF/TOF analysis was carried out for the identification of CH. RESULTS: A series of CH was identified including the main CH groups as HSP90/HATPas_C, HSP70, Cpn60_TCP1, DnaJ, Thioredoxin, TPR, Pro_isomerase, HSP20, ERP29_C, KE2, Prefoldin, DUF704, BAG, GrpE and DcpS. CONCLUSIONS: The ten individual tumor cell lines showed different expression patterns, which are important for the design of CH studies in tumor cell lines. The results can serve as a reference map and form the basis of a concomitant determination of CH by a protein chemical rather than an immunochemical method, independent of antibody availability or specificity.
伴侣蛋白(CH)在肿瘤生物学中发挥着重要作用,但迄今为止尚未有关于其表达模式的系统性研究报道。因此,本研究的目的是提出一种分析方法,用于同时测定人肿瘤细胞系中的几种伴侣蛋白,生成各个细胞系中的表达模式,并寻找肿瘤和非肿瘤细胞系特异性的伴侣蛋白表达情况。将神经母细胞瘤、结直肠癌、卵巢腺癌、骨肉瘤、横纹肌肉瘤、恶性黑色素瘤、肺癌、宫颈癌、乳腺癌、早幼粒细胞白血病的人肿瘤细胞系进行匀浆处理,通过二维凝胶电泳分离蛋白质,对凝胶内的蛋白质进行酶解,并进行基质辅助激光解吸电离飞行时间/串联飞行时间(MALDI-TOF/TOF)分析以鉴定伴侣蛋白。结果:鉴定出一系列伴侣蛋白,包括主要的伴侣蛋白组,如HSP90/HATPas_C、HSP70、Cpn60_TCP1、DnaJ、硫氧还蛋白、TPR、脯氨酸异构酶、HSP20、ERP29_C、KE2、前折叠蛋白、DUF704、BAG、GrpE和DcpS。结论:这十种单独的肿瘤细胞系表现出不同的表达模式,这对于肿瘤细胞系中伴侣蛋白研究的设计很重要。这些结果可作为参考图谱,并构成通过蛋白质化学而非免疫化学方法同时测定伴侣蛋白的基础,该方法独立于抗体的可用性或特异性。