Development of a diagnostic marker and method for pancreatic cancer using secretome and proteome analysis.

• Project/Area Number: 22700904
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Single-year Grants
• Research Field: Tumor diagnosis
• Research Institution: Chiba University
• Principal Investigator: SATOH Mamoru 千葉大学, 医学部・附属病院, 助教 (20401002)
• Co-Investigator(Renkei-kenkyūsha): NOMURA Fumio 千葉大学, 大学院・医学研究院, 教授 (80164739) ; MIYAZAKI Masaru 千葉大学, 大学院・医学研究院, 教授 (70166156) ; YOSHITOMI Hideyuki 千葉大学, 大学院・医学研究院, 助教 (60375631) ; KODERA Yoshio 北里大学, 理学部, 准教授 (60265733) ; SOGAWA Kazuyuki 千葉大学, 医学部・附属病院, 助教 (50436440)
• Research Collaborator: KAWASHIMA Yusuke 北里大学, 理学部, 日本学術振興会特別研究員
• Project Period (FY): 2010 – 2011
• Project Status: Completed (Fiscal Year 2011)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2011: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2010: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
• Keywords: プロテオミクス解析 / プロテオーム / セクレトーム / 安定同位体標識 / 抗がん剤耐性株 / 膵臓がん / 血清

Research Abstract

In this study, we employed a quantitative proteomics technique using stable isotope labeling with amino acids in cell culture (SILAC) followed by mass spectrometry to quantify changes in protein levels between gemcitabine resistant and sensitive pancreatic cancer cells. The 120 proteins were shown more than 2-fold change between the resistant and sensitive cell lines, of which twenty proteins were confirmed by WB. Moreover, 30 proteins were changed more than two times in the comparative analysis of membrane proteins. The siRNA-mediated inhibition of protein expression which was increased in resistant cells increased the cytotoxic effect of gemcitabine. We identified the candidate proteins of diagnosis marker in secretome analysis, the validation of candidate proteins in the serum is now underway.

Research Products

• [Journal Article] Prohibitin in squamous cell carcinoma of the lung : its expression and possible clinical significance (2012)
  • Author(s): Guo F, Hiroshima K, Wu D, Satoh M, Abulazi M, Yoshino I, Tomonaga T, Nomura F, Nakatani Y
  • Journal Title: Human pathology Volume: (in press)
  • Peer Reviewed
• [Journal Article] Plectin promotes migration and invasion of cancer cells and is a novel prognostic marker for head and neck squamous cell carcinoma (2012)
  • Author(s): Katada K, Tomonaga T, Satoh M, Matsushita K, Tonoike Y, Kodera Y, Hanazawa T, Nomura F, Okamoto Y
  • Journal Title: Journal of proteomics Volume: 75(6) Pages: 1803-1815
  • Peer Reviewed
• [Journal Article] Prohibitin in squamous cell carcinoma of the lung : its expression and possible clinical significance (2012)
  • Author(s): Guo F, Hiroshima K, Wu D, Satoh M, Abulazi M, Yoshino I, Tomonaga T, Nomura F, Nakatani Y
  • Journal Title: Human pathology. Volume: (未定)
  • Peer Reviewed
• [Journal Article] Plectin promotes migration and invasion of cancer cells and is a novel prognostic marker for head and neck squamous cell carcinoma (2012)
  • Author(s): Katada K, Tomonaga T, Satoh M, Matsushita K, Tonoike Y, Kodera Y, Hanazawa T, Nomura F, Okamoto Y
  • Journal Title: Journal of proteomics. Volume: 75 Pages: 1803-1815
  • Peer Reviewed
• [Journal Article] The application of a three-step proteome analysis for identification of new biomarkers of pancreatic cancer (2011)
  • Author(s): Abulaizi M, Tomonaga T, Satoh M, Sogawa K, Matsushita K, Kodera Y, Obul J, Takano S, Yoshitomi H, Miyazaki M, Nomura F
  • Journal Title: International journal of proteomics Pages: 628787-628787
  • Peer Reviewed
• [Journal Article] Adsorption of urinary proteins on the conventionally used urine collection analysis and prevention of the adsorption by polymer coating (2011)
  • Author(s): Kiyokawa I, Sogawa K, Ise K, Iida F, Satoh M, Miura T, Kojima R, Katayama K, Nomura F
  • Journal Title: International journal of proteomics Pages: 502845-502845
  • Peer Reviewed
• [Journal Article] Identification of a high molecular weight kininogen fragment as a marker for early gastric cancer by serum proteome analysis (2011)
  • Author(s): Umemura H, Togawa A, Sogawa K, Satoh M, Mogushi K, Nishimura M, Matsushita K, Tanaka H, Takizawa H, Kodera Y, Nomura F
  • Journal Title: J Gastroenterol Volume: 46(5) Pages: 577-85
  • NAID: 10029028943
  • Peer Reviewed
• [Journal Article] dentification of subtilase cytotoxin (SubAB) receptors whose signaling, in association with SubAB-induced BiP cleavage, is responsible for apoptosis in HeLa cel (2011)
  • Author(s): Yahiro K, Satoh M, Morinaga N, Tsutsuki H, Ogura K, Nagasawa S, Nomura F, Moss J, Noda M
  • Journal Title: Infect Immun Volume: 79(2) Pages: 617-27
• [Journal Article] Increased serum levels of pigment epithelium-derived factor by excessive alcohol consumption-detection and identification by a three-step serum proteome analysis (2011)
  • Author(s): Sogawa K, Kodera Y, Satoh M, Kawashima Y, Umemura H, Maruyama K, Takizawa H, Yokosuka O, Nomura F
  • Journal Title: Alcohol Clin Exp Res Volume: 35(2) Pages: 211-7
  • Peer Reviewed
• [Journal Article] Adsorption of urinary proteins on the conventionally used urine collection tubes : possible effects on urinary proteome analysis and prevention of the adsorption by polymer coating (2011)
  • Author(s): Kiyokawa I, Sogawa K, Ise K, Iida F, Satoh M, Miura T, Kojima R, Katayama K, Nomura F
  • Journal Title: International Journal of proteomics. Pages: 502845-502845
  • Peer Reviewed
• [Journal Article] The application of a three-step proteome analysis for identification of new biomarkers of pancreatic cancer (2011)
  • Author(s): Abulaizi M, Tomonaga T, Satoh M, Sogawa K, Matsushita K, Kodera Y, Obul J, Takano S, Yoshitomi H, Miyazaki M, Nomura F
  • Journal Title: International journal of proteomics. Pages: 502845-502845
  • Peer Reviewed
• [Journal Article] Identification of Subtilase Cytotoxin (SubAB) Receptors Whose Signaling, in Association with SubAB-Induced BiP Cleavage, Is Responsible for Apoptosis in HeLa Cells. (2011)
  • Author(s): Yahiro K, Satoh M, Morinaga N, Tsutsuki H, Ogura K, Nagasawa S, Nomura F, Moss J, Noda M.
  • Journal Title: Infect Immun. Volume: 79 Pages: 617-617
  • Peer Reviewed
• [Journal Article] Identification of a high molecular weight kininogen fragment as a marker for early gastric cancer by serum proteome analysis. (2011)
  • Author(s): Umemura H, Togawa A, Sogawa K, Satoh M, Mogushi K, Nishimura M, Matsushita K, Tanaka H, Takizawa H, Kodera Y, Nomura F.
  • Journal Title: J Gastroenterol. Volume: (未定)
  • NAID: 10029028943
  • Peer Reviewed
• [Journal Article] Increased Serum Levels of Pigment Epithelium-Derived Factor by Excessive Alcohol Consumption-Detection and Identification by a Three-Step Serum Proteome Analysis. (2010)
  • Author(s): Sogawa K, Kodera Y, Satoh M, Kawashima Y, Umemura H, Maruyama K, Takizawa H, Yokosuka O, Nomura F
  • Journal Title: Alcohol Clin Exp Res. Volume: 35 Pages: 211-211
  • Peer Reviewed
• [Presentation] 質量分析計を用いた生体資料の藤守定量測定 (2011)
  • Author(s): 佐藤守
  • Organizer: 第8回千葉大学分析センターセミナー(NMR・MS定量分析基礎講座)
  • Place of Presentation: 千葉大学けやき会館3階レセプションホール
  • Year and Date: 2011-09-06
• [Presentation] 質量分析計を用いた生体資料の定量測定 (2011)
  • Author(s): 佐藤守
  • Organizer: 第8回千葉大学分析センターセミナー(NMR・MS定量分析基礎講座)
  • Place of Presentation: 千葉大学けやき会館3階レセプションホール(招待講演)
  • Year and Date: 2011-09-06
• [Presentation] ジェムシタビン耐性ヒト膵癌細胞株のプロテオーム解析 (2011)
  • Author(s): 佐藤守, 高野重紹, 石橋真澄, 吉富秀幸, 西村基, 曽川一幸, 松下一之, 賀川真吾, 荷堂清香, 宮崎勝,野村文夫
  • Organizer: JHUPO第9回大会
  • Place of Presentation: 新潟市朱鷺メッセ
  • Year and Date: 2011-07-28
• [Presentation] 定量的ショットガンプロテオミクスによる歯周疾患マーカーの探索 (2011)
  • Author(s): 土田祥央, 佐藤守, 曽川一幸, 川島祐介, 荷堂清香, 澤井摂, 西村基, 松下一之, 小寺義男, 野村文夫
  • Organizer: JHUPO第9回大会
  • Place of Presentation: 新潟市朱鷺メッセ
  • Year and Date: 2011-07-28
• [Presentation] 放線菌Streptomyces coelicolor A3(2) におけるMsiK. ABC輸送系複合体の探索 (2011)
  • Author(s): 海老瀬広規, 村上聡史, 荷堂清香, 安藤昭一, 齋藤明広
  • Organizer: 第8回21世紀大腸菌研究会
  • Place of Presentation: 南木曽ホテル木曽路
  • Year and Date: 2011-05-18
• [Presentation] 放線菌Streptomyces coelicolor A3(2)におけるMsiK-ABC輸送系複合体の探索 (2011)
  • Author(s): 海老瀬広規, 村上聡史, 佐藤守, 荷堂清香, 安藤昭一, 齋藤明広
  • Organizer: 第8回21世紀大腸菌研究会
  • Place of Presentation: 南木曽 ホテル木曽路
  • Year and Date: 2011-05-18
• [Presentation] プロテオーム解析をもっと身近に (2010)
  • Author(s): 佐藤守
  • Organizer: 資生堂ユーザーズセミナー(リアルLCcafe)
  • Place of Presentation: 東京コンファレンスセンター品川
  • Year and Date: 2010-11-26
• [Presentation] 膵臓癌組織のプロテオーム解析 (2010)
  • Author(s): 佐藤守, 高野重紹, 石橋真澄, 吉富秀幸, 賀川真吾, 宮崎勝, 曽川一幸, 野村文夫
  • Organizer: JHUPO第8回大会
  • Place of Presentation: 東京ベイホテル東急
  • Year and Date: 2010-07-27
• [Patent(Industrial Property Rights)] 免疫測定方法、それに用いるキットおよびそれを用いた癌判定方法 (2010)
  • Inventor(s): 小島良, 野田健太, 宮崎勝, 吉富秀幸, 高野重紹, 野村文夫, 佐藤守, 曽川一幸
  • Industrial Property Rights Holder: 千葉大, 日東紡績
  • Industrial Property Number: 2010-122204
  • Filing Date: 2010-05-28
• [Patent(Industrial Property Rights)] データ収集方法、キット及び腫瘍マーカー (2010)
  • Inventor(s): 小島良, 野田健太, 宮崎勝, 吉富秀幸, 高野重紹, 野村文夫, 佐藤守, 曽川一幸
  • Industrial Property Rights Holder: 千葉大, 日東紡績
  • Industrial Property Number: 2010-129157
  • Filing Date: 2010-05-28
• [Patent(Industrial Property Rights)] 免疫測定方法、それに用いるキットおよびそれを用いた癌判定方法 (2010)
  • Inventor(s): 小島良、野田健太、宮崎勝、吉富秀幸、高野重紹、野村文夫、佐藤守、曽川一幸
  • Industrial Property Rights Holder: 千葉大, 日東紡績
  • Industrial Property Number: 2010-122204
  • Filing Date: 2010-05-28
• [Patent(Industrial Property Rights)] データ収集方法、キット及び腫瘍マーカー (2010)
  • Inventor(s): 小島良、野田健太、宮崎勝、吉富秀幸、高野重紹、野村文夫、佐藤守、曽川一幸
  • Industrial Property Rights Holder: 千葉大, 日東紡績
  • Industrial Property Number: 2010-129157
  • Filing Date: 2010-05-28