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Molecular Medicine

, Volume 21, Issue 1, pp 285–295 | Cite as

Developmental Stage-Specific Hepatocytes Induce Maturation of HepG2 Cells by Rebuilding the Regulatory Circuit

  • Yanning Li
  • Demei Liu
  • Yanhong Zong
  • Jinsheng Qi
  • Bin Li
  • Kun Liu
  • Hui Xiao
Research Article

Abstract

On the basis of their characteristics, we presume that developmental stage-specific hepatocytes should have the ability to induce maturation of hepatoma cells. A regulatory circuit formed by hepatocyte nuclear factor (HNF)-4α, HNF-1α, HNF-6 and the upstream stimulatory factor (USF-1) play a key role in the maturation of embryonic hepatocytes; however, it is unclear whether the regulatory circuit mediates the embryonic induction of hepatoma cell maturation. In this study, 12.5-d to 15.5-d mouse embryonic hepatocytes or their medium were used to coculture or treat HepG2 cells, and the induced maturation was evaluated in vitro and in vivo. In the induced HepG2 cells, the components of the regulatory circuit were detected, their cross-regulation was evaluated and HNF-4α RNA interference was performed. We found that 13.5-d to 14.5-d embryonic hepatocytes could induce HepG2 cell maturation, demonstrated by morphological changes, increased maturation markers and decreased c-Myc and α-fetoprotein (AFP) in vitro. The majority of HepG2 tumors were eliminated by 13.5-d embryonic induction in vivo. All components of the regulatory circuit were upregulated and the binding of HNF-4α, HNF-1α, HNF-6 and USF-1 to their target sites was promoted to rebuild the regulatory circuit in the induced HepG2 cells. Moreover, RNA interference targeting HNF-4α, which is the core of the regulatory circuit, attenuated the induced maturation of HepG2 cells with downregulation of the regulatory circuit. These results revealed that developmental stage-specific hepatocytes could induce the maturation of HepG2 cells by rebuilding the regulatory circuit.

Notes

Acknowledgments

This work was supported by the National Natural Science Foundation of China (81470595) and Hebei Natural Science Foundation (H2012206005).

Supplementary material

10020_2015_2101285_MOESM1_ESM.pdf (978 kb)
Supplementary material, approximately 977 KB.

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Authors and Affiliations

  • Yanning Li
    • 1
  • Demei Liu
    • 2
  • Yanhong Zong
    • 2
  • Jinsheng Qi
    • 2
  • Bin Li
    • 2
  • Kun Liu
    • 2
  • Hui Xiao
    • 2
  1. 1.Department of Molecular Biology, Hebei Key Laboratory of Laboratory AnimalHebei Medical UniversityShijiazhuangChina
  2. 2.Department of Biochemistry, Hebei Key Laboratory of Medical BiotechnologyHebei Medical UniversityShijiazhuangPR China

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