Impairment of liver regeneration by the histone deacetylase inhibitor valproic acid in mice

Background and objective: Liver regeneration is a complex process regulated by a group of genetic and epigenetic factors. A variety of genetic factors have been reported, whereas few investigations have focused on epigenetic regulation during liver regeneration. In the present study, valproic acid (VPA), a histone deacetylase (HDAC) inhibitor, was used to investigate the effect of HDAC on liver regeneration. Methods: VPA was administered via intraperitoneal injection to 2/3 partially hepatectomized mice to detect hepatocyte proliferation during liver regeneration. The mice were sacrificed, and their liver tissues were harvested at sequential time points from 0 to 168 h after treatment. DNA synthesis was detected via a BrdU assay, and cell proliferation was tested using Ki-67. The expressions of cyclin D1, cyclin E, cyclin dependent kinase 2 (CDK2), and CDK4 were detected by Western blot analysis. Chromatin immunoprecipitation (ChIP) assay was used to examine the recruitment of HDACs to the target promoter regions and the expression of the target gene was detected by Western blot. Results: Immunohistochemical analysis showed that cells positive for BrdU and Ki-67 decreased, and the peak of BrdU was delayed in the VPA-administered mice. Consistently, cyclin D1 expression was also delayed. We identified B-myc as a target gene of HDACs by complementary DNA (cDNA) microarray. The expression of B-myc increased in the VPA-administered mice after hepatectomy (PH). The ChIP assay confirmed the presence of HDACs at the B-myc promoter. Conclusions: HDAC activities are essential for liver regeneration. Inhibiting HDAC activities delays liver regeneration and induces liver cell cycle arrest, thereby causing an anti-proliferative effect on liver regeneration.

[1]  Jean S. Campbell,et al.  Liver regeneration. , 2012, Journal of hepatology.

[2]  R. Paro,et al.  Epigenetic reprogramming during tissue regeneration , 2011, FEBS letters.

[3]  N. Fausto,et al.  New concepts in liver regeneration , 2011, Journal of gastroenterology and hepatology.

[4]  M. Ozaki,et al.  Molecular mechanisms of liver regeneration and protection for treatment of liver dysfunction and diseases , 2011, Journal of hepato-biliary-pancreatic sciences.

[5]  R. Merion,et al.  Current Status and Future of Liver Transplantation , 2010, Seminars in liver disease.

[6]  P. Kaldis,et al.  Mammalian cell-cycle regulation: several Cdks, numerous cyclins and diverse compensatory mechanisms , 2009, Oncogene.

[7]  E. Medrano,et al.  HDAC1 Promotes Liver Proliferation in Young Mice via Interactions with C/EBPβ* , 2008, Journal of Biological Chemistry.

[8]  E. Medrano,et al.  HDAC1 Cooperates with C/EBPα in the Inhibition of Liver Proliferation in Old Mice* , 2008, Journal of Biological Chemistry.

[9]  R. Franklin,et al.  Age-dependent epigenetic control of differentiation inhibitors is critical for remyelination efficiency , 2008, Nature Neuroscience.

[10]  Hong Bu,et al.  Clinicopathologic features of pleomorphic hyalinizing angiectatic tumor of soft parts. , 2007, Chinese medical journal.

[11]  M. Avila,et al.  Id2 leaves the chromatin of the E2F4-p130-controlled c-myc promoter during hepatocyte priming for liver regeneration. , 2006, Biochemical Journal.

[12]  R. Burton,et al.  B-myc: N-terminal recognition of myc binding proteins. , 2006, Biochemistry.

[13]  N. Mongan,et al.  Valproic acid, in combination with all-trans retinoic acid and 5-aza-2′-deoxycytidine, restores expression of silenced RARβ2 in breast cancer cells , 2005, Molecular Cancer Therapeutics.

[14]  Gavin Brooks,et al.  Cell Cycle Control , 2004, Methods in Molecular Biology™.

[15]  A. Zimmermann.  Regulation of liver regeneration. , 2004, Nephrology, dialysis, transplantation : official publication of the European Dialysis and Transplant Association - European Renal Association.

[16]  R. Busse,et al.  Valproic acid inhibits angiogenesis in vitro and in vivo. , 2004, Molecular pharmacology.

[17]  J. Meinkoth,et al.  Histone Deacetylase Is a Target of Valproic Acid-Mediated Cellular Differentiation , 2004, Cancer Research.

[18]  H. Koeffler,et al.  Histone Deacetylase Inhibitors Have a Profound Antigrowth Activity in Endometrial Cancer Cells , 2004, Clinical Cancer Research.

[19]  M. Muroni,et al.  Down-regulation of c-myc and Cyclin D1 genes by antisense oligodeoxy nucleotides inhibits the expression of E2F1 and in vitro growth of HepG2 and Morris 5123 liver cancer cells. , 2003, Carcinogenesis.

[20]  Kuang-Hung Cheng,et al.  Histone Deacetylases: Unique Players in Shaping the Epigenetic Histone Code , 2003, Annals of the New York Academy of Sciences.

[21]  J. Albrecht,et al.  Transient expression of cyclin D1 is sufficient to promote hepatocyte replication and liver growth in vivo. , 2001, Cancer research.

[22]  G. Cornwall,et al.  B-Myc, A Proximal Caput Epididymal Protein, Is Dependent on Androgens and Testicular Factors for Expression1 , 2001, Biology of reproduction.

[23]  T. Nakayama,et al.  Participation of histones and histone-modifying enzymes in cell functions through alterations in chromatin structure. , 2001, Journal of biochemistry.

[24]  B. Lutterbach,et al.  B-Myc is preferentially expressed in hormonally-controlled tissues and inhibits cellular proliferation , 2000, Oncogene.

[25]  B M Turner,et al.  Histone acetylation and an epigenetic code. , 2000, BioEssays : news and reviews in molecular, cellular and developmental biology.

[26]  C. Sherr.  The Pezcoller lecture: cancer cell cycles revisited. , 2000, Cancer research.

[27]  P. Gruppuso,et al.  Cell cycle control during liver development in the rat: evidence indicating a role for cyclin D1 posttranscriptional regulation. , 2000, Cell growth & differentiation : the molecular biology journal of the American Association for Cancer Research.

[28]  James M. Roberts,et al.  CDK inhibitors: positive and negative regulators of G1-phase progression. , 1999, Genes & development.

[29]  G. Prendergast,et al.  New Myc-interacting proteins: a second Myc network emerges , 1999, Oncogene.

[30]  L. M. Facchini,et al.  The molecular role of Myc in growth and transformation: recent discoveries lead to new insights , 1998, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[31]  M. Roussel,et al.  Assembly of cyclin D-dependent kinase and titration of p27Kip1 regulated by mitogen-activated protein kinase kinase (MEK1). , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[32]  M. Cole,et al.  Mouse and rat B-myc share amino acid sequence homology with the c-myc transcriptional activator domain and contain a B-myc specific carboxy terminal region. , 1995, Oncogene.

[33]  C. Sherr,et al.  D-type cyclins. , 1995, Trends in biochemical sciences.

[34]  C. Dang,et al.  B-myc inhibits neoplastic transformation and transcriptional activation by c-myc , 1993, Molecular and cellular biology.

[35]  C. Stevens,et al.  Aquaporin 4 and glymphatic flow have central roles in brain fluid homeostasis , 2021, Nature Reviews Neuroscience.

[36]  G. Klein,et al.  Structure and expression of B-myc, a new member of the myc gene family , 1988, Molecular and cellular biology.

[37]  M. Esteller,et al.  Profiling epigenetic alterations in disease. , 2011, Advances in experimental medicine and biology.

[38]  R. Blumenthal,et al.  Introduction--Epiphanies in epigenetics. , 2011, Progress in molecular biology and translational science.

[39]  Xin Chen,et al.  Role of cyclin D1 as a mediator of c-Met- and beta-catenin-induced hepatocarcinogenesis. , 2009, Cancer research.

[40]  A. Pession,et al.  Growth inhibition and proapoptotic activity induction by IIF and valproic acid on RA-resistant leukemia cells. , 2008, Anticancer research.

[41]  H. Willenbring,et al.  A reproducible and well-tolerated method for 2/3 partial hepatectomy in mice , 2008, Nature Protocols.

[42]  David Horn,et al.  Histone deacetylases. , 2008, Advances in experimental medicine and biology.

[43]  P. Kloetzel,et al.  The effects of the histone deacetylase inhibitor valproic acid on cell cycle, growth suppression and apoptosis in multiple myeloma. , 2006, Haematologica.

[44]  M. Serra,et al.  Predictors of morbidity and mortality after the first episode of upper gastrointestinal bleeding in liver cirrhosis. , 2000, Journal of hepatology.

[45]  H. Okayama.  Mammalian Cell Cycle. , 1993 .

[46]  P. L. Altman,et al.  Mouse and rat , 1979 .

[47]  G. Higgins,et al.  Experimental pathology of the liver , 1931 .