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Study on the activity of the signaling pathways regulating hepatocytes from G0 phase into G1 phase during rat liver regeneration

Abstract

Under normal physiological conditions, the majority of hepatocytes are in the functional state (G0 phase). After injury or liver partial hepatectomy (PH), hepatocytes are rapidly activated to divide. To understand the mechanism underlying hepatocyte G0/G1 transition during rat liver regeneration, we used the Rat Genome 230 2.0 Array to determine the expression changes of genes, then searched the GO and NCBI databases for genes associated with the G0/G1 transition, and QIAGEN and KEGG databases for the G0/G1 transition signaling pathways. We used expression profile function (E t ) to calculate the activity level of the known G0/G1 transition signal pathways, and Ingenuity Pathway Analysis 9.0 (IPA) to determine the interactions among these signaling pathways. The results of our study show that the activity of the signaling pathways of HGF, IL-10 mediated by p38MAPK, IL-6 mediated by STAT3, and JAK/STAT mediated by Ras/ERK and STAT3 are significantly increased during the priming phase (2–6 h after PH) of rat liver regeneration. This leads us to conclude that during rat liver regeneration, the HGF, IL-10, IL-6 and JAK/STAT signaling pathways play a major role in promoting hepatocyte G0/G1 transition in the regenerating liver.

Abbreviations

ERK:

extracellular regulated protein kinases

HGF:

hepatocyte growth factor

IL-6:

interleukin-6

IL-10:

interleukin-10

JAK:

Janus kinase

JNK:

c-Jun NH2-terminal kinase

MAPK:

mitogen-activated protein kinase

NF-κB:

nuclear factor κB

STAT3:

signal transducer and activator of transcription

References

  1. Yokoyama, Y., Nagino, M. and Nimura, Y. Mechanisms of hepatic regeneration following portal vein embolization and partial hepatectomy: a review. World J. Surg. 31 (2007) 367–374.

    Article  CAS  PubMed  Google Scholar 

  2. Vondran, F.W., Katenz, E., Schwartlander, R., Morgul, R.S., Haluk, M., Raschzok, N., Gong, X.B., Cheng, X.D., Kehr, D. and Sauer, I.M. Isolation of primary human hepatocytes after partial hepatectomy: criteria for identification of the most promising liver specimen. Artif. Organs 32 (2008) 205–213.

    Article  PubMed  Google Scholar 

  3. Estes, M.D., Do J. and Ahn, C.H. On chip cell separator using magnetic bead-based enrichment and depletion of various surface markers. Biomed. Microdevices 11 (2009) 509–515.

    Article  CAS  PubMed  Google Scholar 

  4. Michalopoulos, G.K. Liver regeneration. J. Cell Physiol. 213 (2007) 286–300.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  5. Kountouras, J., Boura, P. and Lygidakis, N.J. Liver regeneration after hepatectomy. Hepatogastroenterology 48 (2001) 556–562.

    CAS  PubMed  Google Scholar 

  6. Sell, S. The hepatocyte: heperogeneity and plasticity of liver cells. Int. J. Biochem. Cell Biol. 35 (2003) 267–271.

    Article  CAS  PubMed  Google Scholar 

  7. Zimmermann, A. Regulation of liver regeneration. Nephrol. Dial. Transplant. 19 (2004) iv6–iv10.

    Article  CAS  PubMed  Google Scholar 

  8. Fausto, N., Laird, A.D. and Webber, E.M. Liver regeneration. 2. Role of growth factors and cytokines in hepatic regeneration. FASEB J. 9 (1995) 1527–1536.

    CAS  PubMed  Google Scholar 

  9. Cantz, T., Manns, M.P. and Ott, M. Stem cells in liver regeneration and therapy. Cell Tissue Res. 331 (2008) 271–282.

    Article  PubMed Central  PubMed  Google Scholar 

  10. Xu, C.S., Chen, X.G., Chang, C.F., Wang, G.P., Wang, W.B., Zhang, L.X., Zhu, Q.S., Wang, L. and Zhang, F.C. Transcriptional profiles of biliary epithelial cells from rat regenerating liver after partial hepatectomy. Genes Genomics 34 (2012) 245–256.

    Article  CAS  Google Scholar 

  11. Xu, C.S., Yang, Y.J., Yang, J.Y., Chen, X.G. and Wang, G.P. Analysis of the role of the integrin signaling pathway in hepatocytes during rat liver regeneration. Cell. Mol. Biol. Lett. 17 (2012) 274–288.

    Article  CAS  PubMed  Google Scholar 

  12. Michalopeulos, G.K. Liver regeneration. J. Cell Physiol. 213 (2007) 286–300.

    Article  Google Scholar 

  13. Gómez-Lechón, M.J., Guillén, I., Ponsoda, X., Fabra, R., Trullenque, R., Nakamura, T., and Castell, J.V. Cell cycle progression proteins (cyclins), oncogene expression, and signal transduction during the proliferative response of human hepatocytes to hepatocyte growth factor. Hepatology 23 (1996) 1012–1019.

    Article  PubMed  Google Scholar 

  14. Morello, D., Fitzgerald, M.J., Babinet, C. and Fausto, N. C-myc, c-fos and c-jun regulation in the regeneration livers of normal and H-2K/c-myc transgenic mice. Mol. Cell Biol. 10 (1990) 3185–3193.

    CAS  PubMed Central  PubMed  Google Scholar 

  15. Akira, S., Isshiki, H., Sugita, T., Tanabe, O., Kinoshita, S., Nishio, Y., Nakajima, T., Hirano, T. and Kishimoto, T. A nuclear factor for IL-6 expression (NF-IL6) is a member of a C/EBP family. EMBO J. 9 (1990) 1897–1906.

    CAS  PubMed Central  PubMed  Google Scholar 

  16. Zhong, Z., Wen, Z. and Darnell, J.E. Stat3: a STAT family member activated by tyrosine phosphorylation in response to epidermal growth factor and interleukin-6. Science 264 (1994) 95–98.

    Article  CAS  PubMed  Google Scholar 

  17. Trautwein, C., Caelles, C., Vander, G.P., Hunter, T., Karin, M. and Chojkier, M. Transactivation by NF-IL6/LAP is enhanced by phosphorylation of its activation domain. Nature 364 (1993) 544–547.

    Article  CAS  PubMed  Google Scholar 

  18. Cressmann, D.E., Diamond, R.H. and Taub, R. Rapid activation of the Stat3 tran-scription complex in liver regeneration. Hepatology 21 (1995) 1443–1449.

    Article  Google Scholar 

  19. Trautwein, C., Rakemann, T., Niehof, M., Rose-John, S. and Manns, M.P. Acute-phase response factor, increased binding, and target gene transcription during liver regeneration. Gastroenterology 110 (1996) 1854–1862.

    Article  CAS  PubMed  Google Scholar 

  20. Niehof, M., Manns, M.P. and Trautwein, C. CREB controls LAP/C/EBP beta transcription. Mol. Cell Biol. 17 (1997) 3600–3613.

    CAS  PubMed Central  PubMed  Google Scholar 

  21. Fujiyoshi, M. and Ozaki, M. Molecular mechanisms of liver regeneration and protection for treatment of liver dysfunction and diseases. J. Hepatobiliary Pancreat. Sci. 18 (2011) 13–22.

    Article  PubMed  Google Scholar 

  22. Higgins, G.M. and Anderson, R.M. Experimental pathology of the liver: Restoration of the liver of the white rat following partial surgical removal. Arch. Pathol. 12 (1931) 186–202.

    Google Scholar 

  23. Grisham, J.W. Cell types in rat liver cultures: their identification and isolation. Mol. Cell Biochem. 54 (1983) 23–33.

    Google Scholar 

  24. Wang, W.B., Xie, L.F., Wang, W., Wang, L. and Xu C.S. Isolation, purity and identification of hepatocytes in rat normal liver and regenerating liver. Henan Sci. 26 (2008) 1492–1498.

    Google Scholar 

  25. Strober, W. Trypan blue exclusion test of cell viability. Curr. Protoc. Immunol. 2001 May Appendix 3: Appendix 3B.

    Google Scholar 

  26. Twigger, S.N., Smith J., Zuniga-Meyer, A. and Bromberg, S.K. Exploring phenotypic data at the rat genome database. Curr. Protoc. Bioinformatics (2006) Chapter 1, Unit 1.14.

    Google Scholar 

  27. Wang, J.Z., Du, Z., Payattakool, R., Yu, P.S. and Chen, C.F. A new method to measure the semantic similarity of GO terms. Bioinformatics 23 (2007) 1274–1281.

    Article  CAS  PubMed  Google Scholar 

  28. Guo, W., Cai, C., Wang, C., Zhao, L., Wang, L. and Zhang, T. A preliminary analysis of genome structure and composition in Gossypium hirsutum. BMC Genomics 9 (2008) 314.

    Article  PubMed Central  PubMed  Google Scholar 

  29. Xu, C.S., Wang, G.P., Zhang, L.X., Chang, C.F., Zhi, J. and Hao, Y.P. Correlation between liver cancer occurrence and gene expression profiles in rat liver tissue. Genet. Mol. Res. 10 (2011) 3480–3513.

    Article  CAS  PubMed  Google Scholar 

  30. Jiang, C., Xuan, Z., Zhao, F. and Zhang, M.Q. TRED: a transcriptional regulatory element database, new entries and other development. Nucleic Acids Res. 35 (2007) 137–140.

    Article  Google Scholar 

  31. Childress, P.J., Flepcher, R.L. and Perumal, N.B. LymphTF-DB: a database of transcription factors involved in lymphocyte development. Genes Immun. 8 (2007) 360–365.

    Article  CAS  PubMed  Google Scholar 

  32. Wang, G.P. and Xu, C.S. Reference gene selection for real-time RT-PCR in eight kinds of rat regenerating hepatic cells. Mol. Biotechnol. 46 (2010) 49–57.

    Article  PubMed  Google Scholar 

  33. Kost, D.P. and Michalpopulos, G.K. Effect of epidermal growth factor on the expression of protooncogenes c-myc and c-Ha-ras in short-term primary hepatocyte culture. J. Cell. Physiol. 144 (1990) 122–127.

    Article  CAS  PubMed  Google Scholar 

  34. Borowiak, M., Garratt, A.N., Wustefeld, T., Strehle, M., Trautwein, C. and Birchmeier, C. Met provides essential signals for liver regeneration. Proc. Natl. Acad. Sci.USA 101 (2004) 10608–10613.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  35. Okano, J., Shiota, G., Matsumoto, K., Yasui, S., Kurimasa, A., Hisatome, I. Steinbergd, P. and Murawakia, Y. Hepatocyte growth factor exerts a proliferative effect on oval cells through the PI3K/AKT signaling pathway. Biochem. Biophys. Res. Commun. 309 (2003) 298–304.

    Article  CAS  PubMed  Google Scholar 

  36. Stolz, D.B., Mars, W.M., Petersen, B.E., Kim, T.H. and Michalopoulos, G.K. Growth factor signal transduction immediately after two-thirds partial hepatectomy in the rat. Cancers Res. 59 (1999) 3954–3960.

    CAS  Google Scholar 

  37. Boccaccio, C., Ando, M., Tamagnone, L., Bardelli, A., Michieli, P., Battistini, C. and Comoglio, P.M. Induction of epithelial tubules by growth factor HGF depends on the STAT pathway. Nature 391 (1998) 285–288.

    Article  CAS  PubMed  Google Scholar 

  38. Zhang, Y.W., Wang, L.M., Jove, R. and Van de Woude, G.F. Requirement of Stat3 signaling for HGF/SF-Met mediated tumorigenesis. Oncogene 21 (2002) 217–226.

    Article  CAS  PubMed  Google Scholar 

  39. Hirano, T. Interleukin 6 and its receptor: Ten years later. Int. Rev. Immunol. 16 (1998) 249–284.

    Article  CAS  PubMed  Google Scholar 

  40. Lutticken, C., Wegenka, U.M., Yuan, J., Buschmann, J., Schindler, C., Ziemiecki, A., Harpur, A.G., Wilks, A.F., Yasukawa, K., Taga, T., Kishimoto, T., Barbieri, G., Sendtner, M., Pellegrini, S., Heinrich, C. P. and Horn, F. Association of transcription factor APRF and protein kinase Jak1 with the interleukin-6 signaltransducer gp130. Science 263 (1994) 89–92.

    Article  CAS  PubMed  Google Scholar 

  41. Stahl, N., Boulton, T.G., Farruggella, T., Ip. N.Y., Davis, S., Witthuhn, B.A., Quelle, F.W., Silvennoinen, O., Barbieri, G., Pellegrini, S., Ihle, J.N. and Yancopoulos, G.D. Association and activation of Jak Tyk kinases by CNTF-LIF-OSM-IL-6 beta receptor components. Science 263 (1994) 92–95.

    Article  CAS  PubMed  Google Scholar 

  42. Gerhartz, C., Heesel, B., Sasse, J., Hemmann, U., Landgraf, C., Schneider-Mergener, J., Horn, F., Heinrich, P.C. and Graeve, L. Differential activation of acute phase response factor/STAT3 and STAT1 via the cytoplasmic domain of the interleukin 6 signal transducer gp130. I. Definition of a novel phosphotyrosine motif mediating STAT1 activation. J. Biol. Chem. 271 (1996) 12991–12998.

    Article  CAS  PubMed  Google Scholar 

  43. Streetz, K.L., Luedde, T., Manns, M.P. and Trautwein, C. Interleukin 6 and liver regeneration. Gut 47 (2000) 309–312.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  44. Marshall, C. J. Specificity of receptor tyrosine kinase signaling: transient versus sustained extracellular signal-regulated kinase activation. Cell 80 (1995) 179–185.

    Article  CAS  PubMed  Google Scholar 

  45. Cohen, C.B., Ren, R. and Baltimore, D. Modular binding domains in signal transduction proteins. Cell 80 (1995) 237–248.

    Article  CAS  PubMed  Google Scholar 

  46. Hunter, T. Protein kinases and phosphatases: the yin and yang of protein phosphorylation and signaling. Cell 80 (1995) 225–236.

    Article  CAS  PubMed  Google Scholar 

  47. Heldin, C.H. Dimerization of cell surface receptors in signal transduction. Cell 80 (1995) 213–224.

    Article  CAS  PubMed  Google Scholar 

  48. Hill, C.S. and Treisman, R. Transcriptional regulation by extracellular signals: mechanisms and specificity. Cell 80 (1995) 199–212.

    Article  CAS  PubMed  Google Scholar 

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Correspondence to Cunshuan Xu.

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Li, M., Zhou, X., Mei, J. et al. Study on the activity of the signaling pathways regulating hepatocytes from G0 phase into G1 phase during rat liver regeneration. Cell Mol Biol Lett 19, 181–200 (2014). https://doi.org/10.2478/s11658-014-0188-2

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