Weigle Bernd, Fuessel Susanne, Ebner Reinhard, Temme Achim, Schmitz Marc, Schwind Sandra, Kiessling Andrea, Rieger Michael A, Meye Axel, Bachmann Michael, Wirth Manfred P, Rieber E Peter
Institute of Immunology, Medical Faculty, Technical University of Dresden, Germany.
Biochem Biophys Res Commun. 2004 Sep 10;322(1):239-49. doi: 10.1016/j.bbrc.2004.07.106.
The use of molecular targets in novel strategies of tumor treatment largely depends on the identification of proteins with a tumor- or tissue-restricted expression. We identified the novel protein D-GPCR that is selectively overexpressed in human prostate cancer and prostate and belongs to the subfamily of odorant-like orphan G protein-coupled receptors. Quantification of D-GPCR transcripts in different human tissues by real-time PCR demonstrated 27-fold overexpression in prostate compared to skeletal muscle, the organ with second highest transcript numbers in males. Investigation of tumor/normal cDNA pairs obtained from 241 cancer patients including four prostate tumors confirmed the preferential expression in prostate. When comparing the mean transcript level of 15 prostate cancer tissues to their non-tumorous counterparts, D-GPCR was almost 6-fold upregulated. Coupled in vitro transcription and translation of D-GPCR cDNA produced a protein band of approximately 28 kDa. Recombinant, His-tagged protein was expressed in transfected HEK293 cells and gave rise to a 30 kDa band specifically detected by anti-His antibody. These data provide the basis for future studies evaluating the diagnostic potential of D-GPCR and its utility as a novel target in immunotherapy of prostate cancer.
分子靶点在肿瘤治疗新策略中的应用很大程度上取决于对具有肿瘤或组织限制性表达的蛋白质的鉴定。我们鉴定出了新型蛋白质D-GPCR,它在人类前列腺癌和前列腺组织中选择性过表达,属于嗅觉样孤儿G蛋白偶联受体亚家族。通过实时PCR对不同人类组织中D-GPCR转录本进行定量分析,结果显示与骨骼肌相比,前列腺中的转录本表达量高27倍,骨骼肌是男性中转录本数量第二高的器官。对来自241名癌症患者(包括4例前列腺肿瘤)的肿瘤/正常cDNA对进行研究,证实了D-GPCR在前列腺中的优先表达。将15个前列腺癌组织的平均转录水平与其非肿瘤对应组织进行比较时,D-GPCR上调了近6倍。D-GPCR cDNA的体外转录和翻译产生了一条约28 kDa的蛋白带。重组的His标签蛋白在转染的HEK293细胞中表达,并产生了一条可被抗His抗体特异性检测到的30 kDa条带。这些数据为未来评估D-GPCR的诊断潜力及其作为前列腺癌免疫治疗新靶点的效用的研究提供了基础。