Analysis of induced cancer stem-like sphere cells from de-differentiated human hepatocellular carcinoma cell lines to examine the molecular targets for the cancer therapy

• Project/Area Number: 25861192
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Digestive surgery
• Research Institution: Yamaguchi University
• Principal Investigator: TSUNEDOMI Ryouichi 山口大学, 医学(系)研究科(研究院), 助教 (10420514)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000); Fiscal Year 2015: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2014: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2013: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: 癌 / がん幹細胞

Outline of Final Research Achievements

Cancer stem cells (CSCs) are thought to play important roles in therapy-resistance. In this study, we induced cancer stem-like cells from hepatocellular carcinoma (HCC) cell lines using a unique medium, and examined their potential for resistance to anti-cancer drugs. Poorly differentiated HCC derived cell lines efficiently formed spheres of cells, but well-differentiated HCC-derived cells did not. Sphere cells showed increased stemness marker levels. We observed more CD44 variant-positive cells in Sphere cells than in parental cells. The cell viability of Sphere cells was significantly higher than that of parental cells in the presence of several anti-cancer drugs. Sphere cells showed higher ABCG2 and HIF1A mRNA expression and lower ROS production compared to parental cells. Our novel method successfully induced cancer stem-like cells, which possessed chemoresistance that was related to the cell cycle, drug efflux, and ROS.

Research Products

• [Journal Article] Involvement of angiotensin II type 2 receptor (AT2R) signaling in human pancreatic ductal adenocarcinoma (PDAC): a novel AT2R agonist effectively attenuates growth of PDAC grafts in mice. (2015)
  • Author(s): Ishiguro S, Yoshimura K, Tsunedomi R, Oka M, Takao S, Inui M, Kawabata A, Wall T, Magafa V, Cordopatis P, Tzakos AG, Tamura M.
  • Journal Title: Cancer Biol Ther. Volume: 16 Issue: 2 Pages: 307-316
  • DOI: 10.1080/15384047.2014.1002357
  • Peer Reviewed / Open Access
• [Journal Article] Relationship between Polymorphisms and the Efficacy of Cetuximab (2015)
  • Author(s): Yuka Inoue, Shoichi Hazama, Ryouichi Tsunedomi, Sigeru Takeda, Tomio Ueno, Sigeru Yamamoto, Shigefumi Yoshino, Junichi Sakamoto, Hideyuki Mishima, Masaaki Oka, and Hiroaki Nagano
  • Journal Title: Gan To Kagaku Ryoho Volume: 42 Pages: 1310-1312
• [Journal Article] Preoperative serum methylation signature as prognostic tool after curative hepatectomy in patients with hepatocellular carcinoma (2015)
  • Author(s): Shinsuke Kanekiyo, Norio Iizuka, Ryouichi Tsunedomi, Yukio Tokumitsu, Noriaki Hashimoto, Yoshihiro Tokuhisa, Yoshinari Maeda, Michihisa Iida, Kazuhiko Sakamoto, Takao Tamesa, Yusuke Fujita, Shigefumi Yoshino, Shoichi Hazama, Yoshihiko Hamamoto, and Masaaki Oka
  • Journal Title: Anticancer Res Volume: 35 Pages: 997-1007
  • Peer Reviewed
• [Journal Article] A phase ΙI study of five peptides combination with oxaliplatin-based chemotherapy as a first-line therapy for advanced colorectal cancer (FXV study). (2014)
  • Author(s): Hazama S, Nakamura Y, Tanaka H, Hirakawa K, Tahara K, Shimizu R, Ozasa H, Etoh R, Sugiura F, Okuno K, Furuya T, Nishimura T, Sakata K, Yoshimatsu K, Takenouchi H, Tsunedomi R, Inoue Y, Kanekiyo S, Shindo Y, Suzuki N, Yoshino S, Shinozaki H, Kamiya A, Furukawa H, Yamanaka T, Fujita T, Kawakami Y, Oka M.
  • Journal Title: J Transl Med. Volume: 12 Issue: 1 Pages: 108-108
  • DOI: 10.1186/1479-5876-12-108
  • Peer Reviewed / Open Access
• [Journal Article] Predictive Biomarkers for the Outcome of Vaccination of Five Therapeutic Epitope Peptides for Colorectal Cancer (2014)
  • Author(s): Shoichi Hazama, Hiroko Takenouchi, Ryouichi Tsunedomi, Michihisa Iida, Nobuaki Suzuki, Norio Iizuka, Yuka Inoue, Kazuhiko Sakamoto, Mitsuhiro Nakao, Yoshitaro Shindo, Shinsuke Kanekiyo, Yukio Tokumitsu, Kiyoshi Yoshimura, Noriko Maeda, Kazunari Maeda, Yoshinari Maeda, Hiroto Matsui, Shigefumi Yoshino, et al.
  • Journal Title: Anticancer Research Volume: 34 Pages: 4201-4205
  • Peer Reviewed
• [Journal Article] Cancer stem-like sphere cells induced from de-differentiated hepatocellular carcinoma-derived cell lines possess the resistance to anti-cancer drugs (2014)
  • Author(s): Noriaki Hashimoto, Ryouichi Tsunedomi, Kiyoshi Yoshimura, Yusaku Watanabe, Shoichi Hazama, and Masaaki Oka
  • Journal Title: BMC Cancer Volume: 14 Issue: 1 Pages: 722-722
  • DOI: 10.1186/1471-2407-14-722
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] A stem cell medium containing neural stimulating factor induces a pancreatic cancer stem-like cell-enriched population. (2014)
  • Author(s): Watanabe Y, Yoshimura K, Yoshikawa K, Tsunedomi R, Shindo Y, Matsukuma S, Maeda N, Kanekiyo S, Suzuki N, Kuramasu A, Sonoda K, Tamada K, Kobayashi S, Saya H, Hazama S and Oka M
  • Journal Title: Int J Oncol Volume: 45 Issue: 5 Pages: 1857-1866
  • DOI: 10.3892/ijo.2014.2603
  • Peer Reviewed / Open Access
• [Journal Article] Differences in UGT1A1, UGT1A7, and UGT1A9 Polymorphisms between Uzbek and Japanese Populations (2014)
  • Author(s): Hiromichi Maeda, Shoichi Hazama, Abdiev Shavkat, Ken Okamoto, Koji Oba, Junichi Sakamoto, Kenichi Takahashi, Masaki Oka, Daisuke Nakamura, Ryouichi Tsunedomi, Naoko Okayama, Hideyuki Mishima, Michiya Kobayashi
  • Journal Title: Molecular diagnosis & therapy Volume: in press
  • Peer Reviewed
• [Journal Article] Expression of B7-H3, a Potential Factor of Tumor Immune Evasion in Combination with the Number of Regulatory T Cells, Affects Against Recurrence-Free Survival in Breast Cancer Patients (2014)
  • Author(s): Noriko Maeda, Kiyoshi Yoshimura, Sigeru Yamamoto, Atsuo Kuramasu, Moeko Inoue, Nobuaki Suzuki, Yusaku Watanabe, Yoshinari Maeda, Ryoji Kamei, Ryouichi Tsunedomi, Yoshitaro Shindo, Makoto Inui, Koji Tamada, Shigefumi Yoshino, Shoichi Hazama, Masaaki Oka
  • Journal Title: Annals of Surgical Oncology Volume: in press
  • Peer Reviewed
• [Journal Article] A phase I study of combination vaccine treatment of five therapeutic epitope-peptides for metastatic colorectal cancer; safety, immunological response, and clinical outcome (2014)
  • Author(s): Hazama S, Nakamura Y, Takeuchi H, 他全13名
  • Journal Title: J. Transl. Med Volume: 12 (1) Issue: 1 Pages: 63-63
  • DOI: 10.1186/1479-5876-12-63
  • Peer Reviewed
• [Journal Article] UGT1A1*6, 1A7*3, and 1A9*22 genotypes predict severe neutropenia in FOLFIRI-treated metastatic colorectal cancer in two prospective studies in Japan (2013)
  • Author(s): Shoichi Hazama, Ryouichi Tsunedomi, et al.
  • Journal Title: Cancer Science Volume: 104 Issue: 12 Pages: 1662-1669
  • DOI: 10.1111/cas.12283
  • Peer Reviewed
• [Presentation] 肝転移能亢進を示す癌幹細胞様細胞における分子標的の探索 (2015)
  • Author(s): 恒富 亮一、松隈 聰、硲 彰一、岡 正朗、永野 浩昭
  • Organizer: 第26回日本消化器癌発生学会総会
  • Place of Presentation: 米子全日空ホテル (鳥取県)
  • Year and Date: 2015-11-20
• [Presentation] 癌間質におけるmiR221およびmiR222発現は大腸癌の悪性度と相関する (2015)
  • Author(s): 飯田 通久、硲 彰一、恒富 亮一、井上 由佳、徳久 善弘、坂本 和彦、鈴木 伸明、為佐 卓夫、上野 富雄、山本 滋、吉野 茂文、永野 浩昭
  • Organizer: 第26回日本消化器癌発生学会総会
  • Place of Presentation: 米子全日空ホテル (鳥取県)
  • Year and Date: 2015-11-19
• [Presentation] OVEREXPRESSION OF MIRNA221 NAD 222 IN CANCER STROMA IS ASSOCIATED WITH MALIGNANT POTENTIALS IN COLORECTAL CANCER (2015)
  • Author(s): Michihisa Iida, Shoichi Hazama, Ryouichi Tsunedomi, Takao Tamesa, Shigeru Takeda, Tomio Ueno, Shigeru Yamamoto, Shigefumi Yoshino, Yutaka Kawakami, and Hiroaki Nagano
  • Organizer: 23nd United European Gastroenterology Week
  • Place of Presentation: Barcelona, SPAIN
  • Year and Date: 2015-10-26
  • Int'l Joint Research
• [Presentation] miR expression of the tumor might be a predictive biomarker for the efficacy of peptide vaccine for colorectal cancer (2015)
  • Author(s): Shoichi Hazama, Yuka Inoue, Shinsuke Kanekiyo, Yoshitaro Shindo, Nobuaki Suzuki, Takao Tamesa, Ryouichi Tsunedomi, Shigeru Takeda, Tomio Ueno, Shigefumi Yoshino, Yusuke Nakamura, Yutaka Kawakami, and Hiroaki Nagano
  • Organizer: 第74回日本癌学会学術総会
  • Place of Presentation: 名古屋国際会議場 (愛知県)
  • Year and Date: 2015-10-10
• [Presentation] Calreticulin is highly expressed in pancreatic cancer stem like cells (2015)
  • Author(s): Satoshi Matsukuma, Kiyoshi Yoshimura, Yusaku Watanabe, Ryoichi Tsunedomi, Moeko Inoue, Tomoko Kondo-Furuya, Atsunori Oga, Tomio Ueno, Shigefumi Yoshino, Shoichi Hazama, Hiroshi Ito, and Hiroaki Nagano
  • Organizer: 第74回日本癌学会学術総会
  • Place of Presentation: 名古屋国際会議場 (愛知県)
  • Year and Date: 2015-10-08
• [Presentation] Up-regulation of antioxidant ability and HIF1 regulated genes in hepatocellular carcinoma stem-like sphere cells (2015)
  • Author(s): Ryouichi Tsunedomi, Kiyoshi Yoshimura, Shoichi Hazama, and Masaaki Oka
  • Organizer: 第74回日本癌学会学術総会
  • Place of Presentation: 名古屋国際会議場 (愛知県)
  • Year and Date: 2015-10-08
• [Presentation] Overexpression of miR221/222 in cancer stroma are associated with malignant potentials in colorectal cancer (2015)
  • Author(s): Michihisa Iida, Shoichi Hazama, Ryouichi Tsunedomi, Shigeru Takeda, Tomio Ueno, Shigeru Yamamoto, Shigefumi Yoshino, Yutaka Kawakami, and Hiroaki Nagano
  • Organizer: 第74回日本癌学会学術総会
  • Place of Presentation: 名古屋国際会議場 (愛知県)
  • Year and Date: 2015-10-08
• [Presentation] Cancer-stem like細胞の解析による術後肝内再発抑制に向けた分子標的の探索 (2015)
  • Author(s): 恒富 亮一、松隈 聰、吉村 清、坂本 和彦、爲佐 卓夫、硲 彰一、岡 正朗
  • Organizer: 第24回癌病態治療研究会
  • Place of Presentation: 日光千姫物語 (栃木県)
  • Year and Date: 2015-06-26
• [Presentation] 肝細胞癌細胞株より誘導した癌幹細胞様細胞における肝転移能 (2014)
  • Author(s): 恒富 亮一、硲 彰一、岡 正朗
  • Organizer: 第25回日本消化器癌発生学会総会
  • Place of Presentation: ホテル日航福岡 (福岡県)
  • Year and Date: 2014-11-13
• [Presentation] 低分型ヒト肝細胞癌細胞株から誘導培養した癌幹細胞様細胞の解析 (2014)
  • Author(s): 恒富 亮一、吉村 清、橋本 憲輝、渡邊 裕策、松隈 聰、岡 正朗
  • Organizer: 第22回日本消化器関連学会週間 (JDDW2014)
  • Place of Presentation: 神戸国際展示場 (兵庫県)
  • Year and Date: 2014-10-26
• [Presentation] CANCER STEM-LIKE SPHERE CELLS INDUCED FROM DE-DIFFERENTIATED HEPATOCELLULAR CARCINOMA-DERIVED CELL LINES EXERTS LIVER METASTATIC POTENTIAL AND CHEMORESISTANCE (2014)
  • Author(s): Ryouichi Tsunedomi, Kiyoshi Yoshimura, Noriaki Hashimoto, Yusaku Watanabe, Shoichi Hazama, and Masaaki Oka
  • Organizer: 22nd United European Gastroenterology Week (UEGW2014)
  • Place of Presentation: ACV, Vienna, AUSTRIA
  • Year and Date: 2014-10-22
• [Presentation] Induced cancer stem-like sphere cells from de-differentiated human hepatocellular carcinoma cell lines (2014)
  • Author(s): Ryouichi Tsunedomi, Kiyoshi Yoshimura, Shoichi Hazama, and Masaaki Oka
  • Organizer: 第73回日本癌学会学術総会
  • Place of Presentation: パシフィコ横浜 (神奈川県)
  • Year and Date: 2014-09-25
• [Presentation] Biomarkers for immunotherapy: Results from the analysis of a HLA-A-status double-blind, biologically-randomized phase II study of five therapeutic epitope-peptides with oxaliplatin-based chemotherapy as first-line therapy for advanced colorectal cancer (FXV study) (2014)
  • Author(s): Shoichi Hazama, Yusuke Nakamura, Hiroaki Tanaka, Fumiaki Sugiura, Kiyotaka Okuno, Kazuhiko Yoshimatsu, Hiroko Takenouchi, Ryouichi Tsunedomi, Yuka Inoue, Shinsuke Kanekiyo, Yoshitaro Shindo, Kiyoshi Yoshimura, Shigefumi Yoshino, Tomonobu Fujita, Yutaka Kawakami, and Masaaki Oka
  • Organizer: 第18回日本がん免疫学会総会
  • Place of Presentation: ひめぎんホール (愛媛県)
  • Year and Date: 2014-07-31
• [Presentation] 抗癌剤耐性を示すヒト肝細胞癌細胞株から誘導した癌幹細胞様細胞 (2014)
  • Author(s): 恒富亮一、吉村 清、橋本憲輝、渡邊裕策、岡 正朗
  • Organizer: 第69回消化器外科学会総会
  • Place of Presentation: 郡山総合体育館 (福島県)
  • Year and Date: 2014-07-17
• [Presentation] ヒト肝細胞癌細胞株から誘導した癌幹細胞様細胞の解析 (2014)
  • Author(s): 恒富亮一、吉村 清、橋本憲輝、渡邊裕策、松隈 聰、岡 正朗
  • Organizer: 第23回癌病態治療研究会
  • Place of Presentation: ホテルグランヴェール岐山 (岐阜県)
  • Year and Date: 2014-06-12
• [Presentation] 肝細胞癌切除後予後予測マーカーとしてのMinichromosome maintenance complex component 6発現の意義 (2014)
  • Author(s): 飯田道久、飯塚徳男、橋本憲輝、徳光幸生、恒富亮一、前田祥成、徳久善久、坂本和彦、為差卓夫、岡 正朗
  • Organizer: 第114回日本外科学会定期学術集会
  • Place of Presentation: 国立京都国際会館 (京都府)
  • Year and Date: 2014-04-04
• [Presentation] 2つのトランスクリプトーム解析からのHCV関連肝細胞癌予後予測 (2013)
  • Author(s): 恒富 亮一、飯塚 徳男、飯田 通久、筒井 理仁、兼清 信介、坂本 和彦、為佐 卓夫、岡 正朗
  • Organizer: 第113回日本外科学会定期学術集会
  • Place of Presentation: 福岡国際会議場、福岡サンパレス、マリンメッセ福岡(福岡県)
• [Presentation] ヒト膵癌幹細胞における微小周辺環境の探索 (2013)
  • Author(s): 吉村 清、渡邊 裕策、恒富 亮一、新藤 芳太郎、亀井 滝士、前田 訓子、前田 祥成、飯田 道久、鈴木 伸明、上野 富雄、山本 滋、吉野 茂文、硲 彰一、岡 正朗
  • Organizer: 第113回日本外科学会定期学術集会
  • Place of Presentation: 福岡国際会議場、福岡サンパレス、マリンメッセ福岡(福岡県)
• [Presentation] HCV陽性肝細胞癌におけるMCM6発現の意義 (2013)
  • Author(s): 飯田 通久、徳光 幸夫、橋本 憲輝、前田 祥成、徳久 善弘、恒富 亮一、坂本 和彦、為佐 卓夫、飯塚 徳男、岡 正朗
  • Organizer: 第113回日本外科学会定期学術集会
  • Place of Presentation: 福岡国際会議場、福岡サンパレス、マリンメッセ福岡(福岡県)
• [Presentation] ヒト膵癌幹細胞を用いた癌幹細胞richな細胞集団の誘導とその培養液中のサイトカインの検索 (2013)
  • Author(s): 渡邊 裕策、吉村 清、新藤 芳太郎、橋本 憲輝、恒富 亮一、前田 祥成、鈴木 伸明、上野 富雄、吉野 茂文、硲 彰一、岡 正朗
  • Organizer: 第17回日本がん免疫学会総会
  • Place of Presentation: ANAクラウンプラザホテル宇部 (山口県)
• [Presentation] 低分化型肝細胞癌株から誘導された癌幹細胞様細胞の解析 (2013)
  • Author(s): 橋本 憲輝、恒富 亮一、吉村 清、渡邊 裕策、岡 正朗
  • Organizer: 第17回日本がん免疫学会総会
  • Place of Presentation: ANAクラウンプラザホテル宇部 (山口県)
• [Presentation] ヒト膵癌幹細胞からの癌幹細胞の培養による誘導 (2013)
  • Author(s): 吉村 清、渡邊 裕策、倉増 敦朗、井上 萌子、恒富 亮一、新藤 芳太郎、亀井 滝士、前田 訓子、前田 祥成、飯田 道久、鈴木 伸明、上野 富雄、山本 滋、吉野 茂文、硲 彰一、玉田 耕治、岡 正朗
  • Organizer: 第17回日本がん免疫学会総会
  • Place of Presentation: ANAクラウンプラザホテル宇部 (山口県)
• [Presentation] ヒト肝細胞癌細胞株から誘導した癌幹細胞様細胞の抗癌剤耐性 (2013)
  • Author(s): 恒富 亮一、吉村 清、橋本 憲輝、渡邊 裕策、岡 正朗
  • Organizer: 第24回日本消化器癌発生学会総会
  • Place of Presentation: 石川県立音楽堂 (石川県)
• [Presentation] Cancer stem like cells induced from human pancreatic cancer cell lines have epithelial-mesenchymal transition property (2013)
  • Author(s): Yusaku Watanabe, Kiyoshi Yoshimura, Yoshitaro Shindo, Yoshinari Maeda, Ryouichi Tsunedomi, Nobuaki Suzuki, Tomio Ueno, Shigefumi Yoshino, Shoichi Hazama, Masaaki Oka
  • Organizer: 第72回日本癌学会学術総会
  • Place of Presentation: パシフィコ横浜 (神奈川県)
• [Presentation] Chemoresistance of induced cancer-stem cell-like sphere cells from a human hepatocellular carcinoma cell line (2013)
  • Author(s): Ryouichi Tsunedomi, Kiyoshi Yoshimura, Masaaki Oka
  • Organizer: 第72回日本癌学会学術総会
  • Place of Presentation: パシフィコ横浜 (神奈川県)
• [Presentation] Expressions of ULBP1 and NKG2D are related to their clinicopathological outcome in patients with gastric cancer (2013)
  • Author(s): Kiyoshi Yoshimura, Ryoji Kamei, Nobuaki Suzuki, Ryouichi Tsunedomi, Noriko Maeda, Yusaku Watanabe, Yoshinari Maeda, Shigeru Takeda, Tomio Ueno, Shigeru Yamamoto, Shigefumi Yoshino, Shoichi Hazama, Masaaki Oka
  • Organizer: 第72回日本癌学会学術総会
  • Place of Presentation: パシフィコ横浜 (神奈川県)
• [Presentation] Overexpression of MCM6 is associated with poor prognosis of hepatocellular carcinoma (2013)
  • Author(s): Michihisa Iida, Norio Iizuak, Ryouichi Tsunedomi, Kazuhiko Sakamoto, Takao Tamesa, Shoichi Hazama, Masaaki Oka
  • Organizer: 第72回日本癌学会学術総会
  • Place of Presentation: パシフィコ横浜 (神奈川県)
• [Presentation] 抗癌剤耐性を示すヒト肝細胞癌細胞株から誘導した癌幹細胞様細胞 (2013)
  • Author(s): 恒富 亮一、吉村 清、橋本 憲輝、渡邊 裕策、岡 正朗
  • Organizer: 第21回日本消化器関連学会週間
  • Place of Presentation: グランドプリンスホテル新高輪、グランドプリンスホテル高輪、品川プリンスホテル (東京都)
• [Presentation] ヒト膵癌細胞株を用いた癌幹細胞が豊富な細胞集団の誘導 (2013)
  • Author(s): 吉村 清、渡邊 裕策、恒富 亮一、新藤 芳太郎、前田 祥成、松井 洋人、前田 訓子、橋本 憲輝、鈴木 伸明、上野 富雄、山本 滋、吉野 茂文、硲 彰一、岡 正朗
  • Organizer: 第21回日本消化器関連学会週間
  • Place of Presentation: グランドプリンスホテル新高輪、グランドプリンスホテル高輪、品川プリンスホテル (東京都)
• [Presentation] 肝細胞癌におけるMCM6の発現とその意義 (2013)
  • Author(s): 飯田 通久、恒富 亮一、徳久 善弘、坂本 和彦、為佐 卓夫、上野 富雄、吉野 茂文、硲 彰一、岡 正朗
  • Organizer: 第21回日本消化器関連学会週間
  • Place of Presentation: グランドプリンスホテル新高輪、グランドプリンスホテル高輪、品川プリンスホテル (東京都)
• [Presentation] CANCER-STEM CELL-LIKE SPHERE CELLS INDUCED FROM A CELL LINE DERIVED FROM POORLY-DIFFERENTIATED HEPATOCELLULAR CARCINOMA EXERTS LIVER METASTATIC POTENTIAL AND CHEMORESISTANCE (2013)
  • Author(s): Ryouichi Tsunedomi, Kiyoshi Yoshimura, Noriaki Hashimoto, Yusaku Watanabe, Masaaki Oka
  • Organizer: 21st United European Gastroenterology Week
  • Place of Presentation: ICC Berlin, Berlin, The Federal Republic of GERMANY
• [Patent(Industrial Property Rights)] 肝細胞がんの転移性再発リスクの予測方法 (2015)
  • Inventor(s): 岡 正朗、硲 彰一、恒富 亮一
  • Industrial Property Rights Holder: 山口大学
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2015-016586
  • Filing Date: 2015-01-30