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

• Project/Area Number: 17390439
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Urology
• Research Institution: Kyoto University
• Principal Investigator: NAKAMURA Eijiro Kyoto University, Graduate school of medicine, associate professor (90293878)
• Co-Investigator(Kenkyū-buntansha): OGAWA Osamu Kyoto university, Graduate school of medicine, professor (90260611) ; KAMOTO Toshiyuki Kyoto university, Graduate school of medicine, associate professor (00281098) ; NISHIYAMA Hiroyuki Kyoto university, Graduate school of medicine, associate professor (20324642) ; 高橋 毅 京都大学, 医学研究科, 助手 (30362487)
• Project Period (FY): 2005 – 2007
• Project Status: Completed (Fiscal Year 2007)
• Budget Amount: ¥11,740,000 (Direct Cost: ¥11,200,000、Indirect Cost: ¥540,000); Fiscal Year 2007: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000); Fiscal Year 2006: ¥4,900,000 (Direct Cost: ¥4,900,000); Fiscal Year 2005: ¥4,500,000 (Direct Cost: ¥4,500,000)
• Keywords: 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

Research Abstract

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.

Research Products

• [Journal Article] Secreted CXCL1 is a potential Mediator and Marker of the Tumor Invasion of Bladder Cancer (2008)
  • Author(s): Kawanishi H, Nishiyama H, Kamoto T, Nakamura E, Ogawa O.
  • Journal Title: Clin Cancer Res 14 Pages: 2579-2587
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Secreted CXCL1 Is a Potential Mediator and Marker of the Tumor Invasion of Bladder Cancer (2008)
  • Author(s): Kwanishi, H., Matsui, Y., Ito, M., Watanabe, J., Takahashi, T., Nishizawa, K., Nishiyama, H., Kamoto, T., Mikami, Y., Tanaka, Y., Jung, G., Akiyama, H., Nobumasa, H., Guilford, P., Reeve, A., Okuno, Y., Tsujimoto, G., Nakamura, E., Ogawa, O
  • Journal Title: Clin Cancer Res 14 Pages: 2579-87
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Secreted CXCL1 is a potential Mediator and Marker of the Tumor Invasion of Bladder Cancer (2008)
  • Author(s): Kawanishi H, Nishiyama H, Kamoto T, Nakamura E, Ogawa O.
  • Journal Title: Clin Cancer Res (in press)
  • Peer Reviewed
• [Journal Article] STAT3 polymorphism predicts interferon-alfa response in patients with metastatic renal cell carcinoma. (2007)
  • Author(s): Ito N, Nakamura E, Ogawa O.
  • Journal Title: J Clin Oncol 25 Pages: 2785-2791
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] STAT3 polymorphism predicts the interferon-response in patients with metastatic renal cell carcinoma (2007)
  • Author(s): N., Ito, M., Eto, E., Nakamura, A., Takahashi, T., Tsukamoto, H., Toma, H., Nakazawa, Y., Hirao, H., Uemura, S., Kagawa, H., Kanayama, Y., Nose, N., Kinukawa, T., Nakamura, N., Jinnai, T., Seki, M., Takamatsu, Y., Masui, S., Naito, Osamu, Ogawa
  • Journal Title: J Clin Onco 125 Pages: 2785-2791
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Clusterin is a secreted marker for a hypoxia-inducible factor-independent function of Von Hippel-Lindau tumor suppressor protein. (2006)
  • Author(s): Nakamura E, Kamoto T, Ogawa O, Kaelin WG.
  • Journal Title: Am J Pathol. 168 Pages: 574-584
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Kaelin. Clusterin is a secreted marker for a hypoxia-inducible factor-independent function of Von Hippel-Lindau tumor suppressor protein (2006)
  • Author(s): Nakamura, P., Abeu-e-Lima, Y., Awakura, T., Inoue, T., Kamoto, O., Ogawa, H., Kotani, T., Manabe, G., Zhang, K., Kondo, V., Nose, WG
  • Journal Title: Am J Pathol 168 Pages: 574-584
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Clusterin is a Secreted Marker for a HIF-independent Function of the VHL Tumor Suppressor Protein. (2006)
  • Author(s): Nakamura E, Abreu-e-Lima P, Awakura Y, Inoue T, Kamoto T, Ogawa O, Kotani H, Manabe T, Zhang GJ, Kondo K, Nose V, Kaelin WG Jr.
  • Journal Title: Am J Pathol 168(2): Pages: 574-584
• [Journal Article] Neuronal Apoptosis linked to EglN3 Proly1 Hydroxylase and Familial Pheochromocytoma Genes : Developmental Culling and Cancer. (2006)
  • Author(s): S Lee^*, E Nakamura^*, H Yang, W Wei, MS Linggi, MP.Sajan, RV Farese, Rs Freeman, BD Carter, WG Kaelin, S Schlisio
  • Journal Title: Cancer Cell 8 Pages: 155-167
• [Journal Article] Neuronal Apoptosis linked to EglN3 Prolyl Hydroxylase and Familial Pheochromocytoma Genes: Developmental Culling and Cancer. (2005)
  • Author(s): Lee S, Nakamura E, Yang H, Wei W, Linggi MS, Sajan MP, Farese RV, Freeman RS, Carter BD, Kaelin WG, and Schlisio S.
  • Journal Title: Cancer Cell 8 Pages: 155-167
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Neuronal Apoptosis linked to EgIN3 Prolyl Hydroxylase and Familial Pheochromocytoma Genes : Developmental Culling and Cancer. (Co-First Author) (2005)
  • Author(s): S., Lee, E., Nakamura, H., Yang, W., Wei, MS., Linggi, MP., Sajan, RV., Farese, RS., Freeman, BD., Carter, WG., Kaelin, S., Schlisio
  • Journal Title: Cancer Cells 8 Pages: 155-167
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Antiandrogen bicalutamide promotes tumor growth in a novel androgen dependent prostate cancer xenograft model derived from a bicalutamide treated patient. (2005)
  • Author(s): T.Yoshida, H.Kinoshita, T.Segawa, E.Nakamura, T.Inoue, Y.Shimizu, T.Kamoto, O.Ogawa
  • Journal Title: Cancer Res. 65(21) Pages: 9611-9616
• [Journal Article] Multiple roles of Rbx1 in the VBC-Cul2 ubiquitin ligase complex. (2005)
  • Author(s): Megumi Y, Miyauchi Y, Sakurai H, Nobeyama H, Lorick K, Nakamura E, Chiba T, Tanaka K, Weissman AM, Kirisako T, Ogawa O, Iwai K
  • Journal Title: Genes Cells 10(7) Pages: 679-691
• [Journal Article] Monthly paclitaxel and carboplatin with oral estramustine phosphate in patients with hormone refractory prostate cancer. (2005)
  • Author(s): T Segawa, T Kamoto, E Nakamura, O Ogawa
  • Journal Title: Int J Clin.Oncology 10: Pages: 333-337
• [Journal Article] The expression of NOV and WT1 in renal cell carcinoma : a quantitative reverse transcriptase-polymerase chain reaction analysis. (2005)
  • Author(s): Niu Z, Ito M, Awakura Y, Takahashi T, Nakamura E, Ito N, Ogawa O.
  • Journal Title: J Urol 174: Pages: 1460-1462
• [Presentation] Cell culture proteome identifies CXCL1 as a secreted marker and mediator for the tumor invasion of bladder cancer. (2007)
  • Author(s): 川西 博晃、中村 英二郎、西山 博之、賀本 敏行、小川 修
  • Organizer: 2007米国泌尿器科学会総会
  • Place of Presentation: アメリカ、アナハイム
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Secreted CXCL1 is a Mediator and Marker of the Tumor Invasion of Bladder Cancer (2007)
  • Author(s): Kwanishi, H., Nakamura, E., Nishiyama, H., Kamoto, T., Ogawa, O
  • Organizer: 2007 annual congress meeting of AUA
  • Place of Presentation: Anahaim, USA
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Identification of serum biomarkers of renal cell carcinoma by proteomic analysis (2006)
  • Author(s): Yamasaki, T., Nakamura, E., Kwanishi, H., Awakura, Y., Kamba, T., Ito, N., Kamoto, T., Ogawa, O
  • Organizer: 65th annual meeting of Japanese Cancer Association
  • Place of Presentation: yokohama
  • Year and Date: 2006-09-29
  • Description: 「研究成果報告書概要(欧文)」より