Identification and validation of tumor biomarkers for urogenital cancer using proteomic analysis of cell culture supernatant

使用细胞培养上清液的蛋白质组学分析鉴定和验证泌尿生殖癌的肿瘤生物标志物

• 批准号: 17390439
• 负责人: NAKAMURA Eijiro
• 金额: $ 7.51万
• 依托单位: Kyoto University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-17390439/
• 关键词: urogenital cancer; tumor maker; shotgun proteomics; cell culture supernatant; 泌尿生殖器癌; 診断マーカー; プロテオミクス; Clusterin; 腎細胞癌; EGLN3; 副腎褐色細胞腫

New proteomic approach for the identification of proteins that are potentially involved in the tumor invasion of bladder cancer.We searched for the candidate proteins by comparing the profiles of secreted proteins among the poorly invasive human bladder carcinoma cell line and the highly invasive cell line. The proteins isolated from cell culture supernatants were identified by shotgun proteomics. We found that CXCL1 modulates the invasive abilities of bladder cancer cells and. this chemokine maybe a potential candidate of urinary biomarker for invasive bladder cancer and a possible therapeutic target for preventing tumor invasion.New proteomic approach for the identification and validation of novel serum tumor makers for renal cell carcinoma.We established a new method fir analyzing the secreted proteins in cell culture supernatants by using serum-free media and compared the proteome among 786-O RCC (VHL-/-) subclones stably transfected with either a plasmid encoding wild-type pVHL or an empty expression plasmid. By using this method, we identified three different VHL regulated secreted proteins (PAI-1,IGFBP-3 and clusterin). IGFBP-3 is elevated significantly in the serum from patients with ccRCC comparing with that from normal controls. On the other hand, we had done the preliminary analysis of "serum proteome" of 4 patients with sporadic clear cell RCC before and after the nephrectomy by using novel multidimensional shotgun proteomics. A total of 6 proteins were identified as candidate markers that were detected before the operation, but disappeared after the procedure. Two of them were biddy expressed in tumor specimens comparing with that of normal adjacent normal tissue. Now we perform further validation of these candidate proteins.

新的蛋白质组学方法用于鉴定可能参与膀胱癌肿瘤侵袭的蛋白质。我们通过比较低侵袭性和高侵袭性人膀胱癌细胞系的分泌蛋白质谱来寻找候选蛋白质。通过鸟枪蛋白质组学鉴定从细胞培养上清液中分离的蛋白质。我们发现CXCL 1调节膀胱癌细胞的侵袭能力。本研究建立了一种新的蛋白质组学方法，对786-O RCC（VHL-/-）细胞培养上清液中分泌蛋白进行了分析，并对786-O RCC（VHL-/-）细胞的蛋白质组进行了比较，结果表明，786-O RCC（VHL-/-）用编码野生型pVHL的质粒或空表达质粒稳定转染的亚克隆。利用这种方法，我们鉴定了三种不同的VHL调节分泌蛋白（派-1，IGFBP-3和clusterin）。与正常对照组相比，ccRCC患者血清中IGFBP-3水平显著升高。另一方面，我们采用新的多维鸟枪蛋白质组学技术对4例散发性透明细胞肾细胞癌患者手术前后的血清蛋白质组进行了初步分析。共有6种蛋白被鉴定为候选标记物，这些标记物在手术前被检测到，但在手术后消失。其中2个在肿瘤组织中的表达高于癌旁正常组织。现在我们对这些候选蛋白进行进一步的验证。

项目成果

Cell culture proteome identifies CXCL1 as a secreted marker and mediator for the tumor invasion of bladder cancer

细胞培养蛋白质组将 CXCL1 鉴定为膀胱癌肿瘤侵袭的分泌标记物和介质

• 发表时间: 2007
• 作者: Kawanishi H;Ogawa O;et. al.
• 通讯作者: et. al.

Neuronal Apoptosis linked to EglN3 Proly1 Hydroxylase and Familial Pheochromocytoma Genes : Developmental Culling and Cancer.

与 EglN3 Proly1 羟化酶和家族性嗜铬细胞瘤基因相关的神经元凋亡：发育剔除和癌症。

• 发表时间: 2006
• 期刊: Cancer Cell 8
• 作者: S Lee^*;E Nakamura^*;H Yang;W Wei;MS Linggi;MP.Sajan;RV Farese;Rs Freeman;BD Carter;WG Kaelin;S Schlisio
• 通讯作者: S Schlisio

STAT3 polymorphism predicts interferon-alfa response in patients with metastatic renal cell carcinoma

• DOI: 10.1200/jco.2006.09.8897
• 发表时间: 2007-07-01
• 期刊: JOURNAL OF CLINICAL ONCOLOGY
• 影响因子: 45.3
• 作者: Ito, Noriyuki;Eto, Masatoshi;Ogawa, Osamu
• 通讯作者: Ogawa, Osamu

Monthly paclitaxel and carboplatin with oral estramustine phosphate in patients with hormone-refractory prostate cancer.

激素难治性前列腺癌患者每月接受紫杉醇和卡铂联合口服磷酸雌莫司汀。

• 发表时间: 2005
• 期刊: Int.J.Clin.Oncol. 10
• 作者: Segawa;T.;Yamamoto;S.et al.
• 通讯作者: S.et al.

STAT3 polymorphism predicts the interferon- response in patients with metastatic renal cell carcinoma.

STAT3 多态性预测转移性肾细胞癌患者的干扰素反应。

• 发表时间: 2007
• 期刊: J. Clin. Oncol. (In press)
• 作者: Iwasaki M;Hayashi Y;Kamibayashi T;Mashikmo T.;Hayashi Y.;林 行雄;Yokomizo A. et al.;Ito N.et al.
• 通讯作者: Ito N.et al.