ORGANOTYPIC LIVER CULTURES AND HEPATOCARCINOGENESIS

Summary

Principal Investigator: ROBERT FLOYD
Affiliation: University of Oklahoma Health Sciences Center
Country: USA
Abstract: Hepatocarcinogenesis is a complex process requiring cell-to-cell interactions within the three-dimensional tissue architecture that constitutes the mammalian liver. We seek to enhance our abilities to investigate hepatocarcinogenesis by extending the process to the culture dish as much as possible. We have developed an organotypic rat liver model in which viable tissue has been maintained in culture for longer than 90 days. The overall objective for this application is to further develop this organotypic culture model for rat liver (and in the future, for human liver tumor tissue, as well), and to demonstrate its usefulness for investigating choline-deficient hepatocarcinogenesis in a rat model. Toward this end, we propose three Specific Aims: (1) For our organotypic rat liver culture model, we will analyze and test all medium components until we arrive at a defined medium that provides long periods of viability as judged by dual vital staining, by gene expression patterns revealed by DNA arrays, by metabolic capabilities revealed by enzyme amounts and activities. (2) We will further characterize CWSV-1 cells, which form islands of tissue in culture that approximate normal liver in pattern of organization, especially as to their ability to give rise to more than one liver cell type and their progression to tumorigenic cells in choline-deficient medium. (3) We will compare and contrast these two culture models in Aims (1) and (2) with respect to the ability of PBN and 4-OH-PBN to increase apoptosis and inhibit tumor development. Ultimately, we seek an organotypic culture model in which the full process of carcinogenesis from tumor initiation through progression to full metastatic cancer can be manipulated and cures tested.
Funding Period: 1999-07-01 - 2006-01-31
more information: NIH RePORT

Top Publications

  1. ncbi Sensitivity of protein tyrosine phosphatase activity to the redox environment, cytochrome C, and microperoxidase
    Scott J Salsman
    Free Radical Biology and Aging Research Program, Oklahoma Medical Research Foundation, Oklahoma City, OK 73104, USA
    Antioxid Redox Signal 7:1078-88. 2005
  2. ncbi Serendipitous findings while researching oxygen free radicals
    Robert A Floyd
    Experimental Therapeutics Research Program, Oklahoma Medical Research Foundation, Oklahoma City, OK 73170, USA
    Free Radic Biol Med 46:1004-13. 2009
  3. ncbi In vivo MRS assessment of altered fatty acyl unsaturation in liver tumor formation of a TGF alpha/c-myc transgenic mouse model
    J Griffitts
    Advanced Magnetic Resonance Center, Oklahoma Medical Research Foundation, Oklahoma City, OK 73104, USA
    J Lipid Res 50:611-22. 2009
  4. ncbi Nitrones as therapeutics
    Robert A Floyd
    Experimental Therapeutics Research Program, Advanced Magnetic Resonance Center, Oklahoma Medical Research Foundation, Oklahoma City, OK 73104, USA
    Free Radic Biol Med 45:1361-74. 2008
  5. ncbi Modulation of Fas-FasL related apoptosis by PBN in the early phases of choline deficient diet-mediated hepatocarcinogenesis in rats
    Yoshiko Inoue
    Free Radical Biology and Aging Research Program, Oklahoma City, OK 73104, USA
    Free Radic Res 41:972-80. 2007
  6. ncbi Expression of inducible nitric oxide (NO) synthase but not prevention by its gene ablation of hepatocarcinogenesis with fibrosis caused by a choline-deficient, L-amino acid-defined diet in rats and mice
    Ayumi Denda
    Department of Molecular Pathology, Nara Medical University, 840 Shijo Cho, Kashihara, Nara 634 8521, Japan
    Nitric Oxide 16:164-76. 2007
  7. ncbi DNA cytosine methylation profile in various cancer-related genes is altered in cultured rat hepatocyte cell lines as compared with primary hepatocytes
    Kiyoshi Asada
    Third Department of Internal Medicine, Nara Medical University, Kashihara, Nara 634 8521, Japan
    Oncol Rep 15:1241-8. 2006
  8. ncbi Nitrones as therapeutics in age-related diseases
    Robert A Floyd
    Oklahoma Medical Research Foundation, Oklahoma City, OK 73104, USA
    Aging Cell 5:51-7. 2006
  9. ncbi Nitric oxide production by primary liver cells isolated from amino acid diet-fed rats
    Yashige Kotake
    Oklahoma Medical Research Foundation, Free Radical Biology and Aging Research Program, Oklahoma City, OK 73104, USA
    Methods Enzymol 396:535-41. 2005
  10. ncbi Temporal correlation of pathology and DNA damage with gene expression in a choline-deficient model of rat liver injury
    Christine L Powell
    Curriculum in Toxicology, School of Medicine, University of North Carolina, Chapel Hill, NC 27599-7431, USA
    Hepatology 42:1137-47. 2005

Scientific Experts

  • Robert A Floyd
  • Fumiyuki Uematsu
  • Yashige Kotake
  • Ayumi Denda
  • Dai Nakae
  • J Griffitts
  • Yoshiko Inoue
  • Hideki Kishida
  • Kiyoshi Asada
  • Scott J Salsman
  • Christine L Powell
  • G E Reid
  • R A Towner
  • D Saunders
  • Y Tesiram
  • Osamu Kusuoka
  • Debra Saunders
  • Hiroki Kuniyasu
  • Nataliya Smith
  • Yoichi Konishi
  • Taketoshi Asanuma
  • Rheal A Towner
  • Jenny Oblander
  • Nao Murata
  • Eiji Kita
  • Kazutoshi Tamura
  • Fumiko Nishikawa
  • Masahiro Tsutsumi
  • Wakashi Kitayama
  • Hitoshi Yoshiji
  • Rumiko Asada
  • Hiroshi Fukui
  • Edward K Lobenhofer
  • Blair U Bradford
  • Kenneth Hensley
  • Ivan Rusyn
  • Oksana Kosyk
  • Joel S Parker

Detail Information

Publications12

  1. ncbi Sensitivity of protein tyrosine phosphatase activity to the redox environment, cytochrome C, and microperoxidase
    Scott J Salsman
    Free Radical Biology and Aging Research Program, Oklahoma Medical Research Foundation, Oklahoma City, OK 73104, USA
    Antioxid Redox Signal 7:1078-88. 2005
    ..Taken together, these data suggest that not only H2O2, but also a variety of redox-active metabolites and hemes can oxidatively inactivate PTPs with potentially profound implications for signal transduction...
  2. ncbi Serendipitous findings while researching oxygen free radicals
    Robert A Floyd
    Experimental Therapeutics Research Program, Oklahoma Medical Research Foundation, Oklahoma City, OK 73170, USA
    Free Radic Biol Med 46:1004-13. 2009
    ..More recently we discovered that PBN nitrones have potent anti-cancer activity and are active in preventing hearing loss caused by acute acoustical trauma...
  3. ncbi In vivo MRS assessment of altered fatty acyl unsaturation in liver tumor formation of a TGF alpha/c-myc transgenic mouse model
    J Griffitts
    Advanced Magnetic Resonance Center, Oklahoma Medical Research Foundation, Oklahoma City, OK 73104, USA
    J Lipid Res 50:611-22. 2009
    ....
  4. ncbi Nitrones as therapeutics
    Robert A Floyd
    Experimental Therapeutics Research Program, Advanced Magnetic Resonance Center, Oklahoma Medical Research Foundation, Oklahoma City, OK 73104, USA
    Free Radic Biol Med 45:1361-74. 2008
    ....
  5. ncbi Modulation of Fas-FasL related apoptosis by PBN in the early phases of choline deficient diet-mediated hepatocarcinogenesis in rats
    Yoshiko Inoue
    Free Radical Biology and Aging Research Program, Oklahoma City, OK 73104, USA
    Free Radic Res 41:972-80. 2007
    ..The CD diet model is also considered as a model for non-alcoholic steatohepatitis (NASH) in humans and early expression of Fas could also be a good index of the progression of NASH...
  6. ncbi Expression of inducible nitric oxide (NO) synthase but not prevention by its gene ablation of hepatocarcinogenesis with fibrosis caused by a choline-deficient, L-amino acid-defined diet in rats and mice
    Ayumi Denda
    Department of Molecular Pathology, Nara Medical University, 840 Shijo Cho, Kashihara, Nara 634 8521, Japan
    Nitric Oxide 16:164-76. 2007
    ..Results suggested no major determinant roles of the expressed iNOS in the development of liver tumors caused by the CDAA diet...
  7. ncbi DNA cytosine methylation profile in various cancer-related genes is altered in cultured rat hepatocyte cell lines as compared with primary hepatocytes
    Kiyoshi Asada
    Third Department of Internal Medicine, Nara Medical University, Kashihara, Nara 634 8521, Japan
    Oncol Rep 15:1241-8. 2006
    ..This study suggests that a cautious approach is required when cell lines are utilized to study methylation-related carcinogenic, metastatic or tumoricidal mechanisms...
  8. ncbi Nitrones as therapeutics in age-related diseases
    Robert A Floyd
    Oklahoma Medical Research Foundation, Oklahoma City, OK 73104, USA
    Aging Cell 5:51-7. 2006
    ....
  9. ncbi Nitric oxide production by primary liver cells isolated from amino acid diet-fed rats
    Yashige Kotake
    Oklahoma Medical Research Foundation, Free Radical Biology and Aging Research Program, Oklahoma City, OK 73104, USA
    Methods Enzymol 396:535-41. 2005
    ..L-arginine or L-glutamine (material for L-arginine biosynthesis) enriched diet showed the same NO augmentation as in AA diet. It is speculated that a high content of free L-arginine in AA diet may have caused enhanced NO production...
  10. ncbi Temporal correlation of pathology and DNA damage with gene expression in a choline-deficient model of rat liver injury
    Christine L Powell
    Curriculum in Toxicology, School of Medicine, University of North Carolina, Chapel Hill, NC 27599-7431, USA
    Hepatology 42:1137-47. 2005
    ....
  11. ncbi Distinct patterns of gene expression in hepatocellular carcinomas and adjacent non-cancerous, cirrhotic liver tissues in rats fed a choline-deficient, L-amino acid-defined diet
    Fumiyuki Uematsu
    Department of Pathology, Sasaki Institute, Sasaki Foundation, Chiyoda ku, Tokyo, Japan
    Cancer Sci 96:414-24. 2005
    ....
  12. ncbi Anti-cancer activity of nitrones in the Apc(Min/+) model of colorectal cancer
    Robert A Floyd
    Experimental Therapeutics Research Program, Oklahoma Medical Research Foundation, Oklahoma City, OK 73104, USA
    Free Radic Res 44:108-17. 2010
    ..PBN has significant anti-cancer activity in this model of intestinal neoplasia...