Transduction of immortalized human hepatocytes with p21 to enhance differentiated phenotypes

T. Kunieda, N. Kobayashi, Masakiyo Sakaguchi, T. Okitsu, T. Totsugawa, T. Watanabe, T. Matsumura, M. Maruyama, H. Noguchi, M. Takesue, N. Shibata, K. Ohmoto, Toshiyoshi Fujiwara, S. Yamamoto, N. Tanaka

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

We previously constructed an immortal human hepatocyte line NKNT-3 with a simian virus 40 T antigen (SV40T) to develop cell-based biological therapies. p21 is a molecule that regulates the transition from the G1 phase to the S phase of the cell cycle. Investigators have demonstrated that overexpression of p21 induces differentiation in various cell lines. In the current study we examined the effect of p21 on differentiated phenotypes of SV40T-immortalized NKNT-3 cells. A replication-deficient adenovirus vector expressing a human wild-type p21 cDNA under the control of the CMV promoter (Ad5CMVp21) and a human wild-type p21 protein fused to the protein transduction domain from the human immunodeficiency virus (HIV) TAT protein (TAT/p21) were utilized to achieve efficient delivery of p21 into NKNT-3 cells. Morphological alterations, cell cycle progression, and expression of albumin and p-450 associated enzymes (CYPs) 3A4 and 2C9 were evaluated in NKNT-3 cells treated with Ad5CMVp21 and TAT/p21. Efficient adenovirus-based p21 transfer and TAT-mediated p21 protein delivery were confirmed in NKNT-3 cells in an immunofluorescence study and Western blotting analysis. Transduction of NKNT-3 cells with p21 predominantly arrested the cell cycle at the G1 checkpoint, resulting in differentiated hepatic phenotypes in morphology and improvement in protein expression of albumin, CYP 3A4, and CYP C29. We here show that exogenous expression of p21 augments cellular differentiation in immortalized human NKNT-3 cells.

Original languageEnglish
Pages (from-to)421-428
Number of pages8
JournalCell Transplantation
Volume11
Issue number5
Publication statusPublished - 2002

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Hepatocytes
Proteins
Phenotype
Viruses
Cells
Antigens
Simian virus 40
Viral Tumor Antigens
Adenoviridae
Albumins
Cell Cycle
Human Immunodeficiency Virus Proteins
G1 Phase Cell Cycle Checkpoints
Biological Therapy
G1 Phase
Enzymes
S Phase
Fluorescent Antibody Technique
Molecules
Complementary DNA

Keywords

  • Immortalized hepatocytes
  • p21
  • Protein transduction

ASJC Scopus subject areas

  • Cell Biology
  • Transplantation

Cite this

Kunieda, T., Kobayashi, N., Sakaguchi, M., Okitsu, T., Totsugawa, T., Watanabe, T., ... Tanaka, N. (2002). Transduction of immortalized human hepatocytes with p21 to enhance differentiated phenotypes. Cell Transplantation, 11(5), 421-428.

Transduction of immortalized human hepatocytes with p21 to enhance differentiated phenotypes. / Kunieda, T.; Kobayashi, N.; Sakaguchi, Masakiyo; Okitsu, T.; Totsugawa, T.; Watanabe, T.; Matsumura, T.; Maruyama, M.; Noguchi, H.; Takesue, M.; Shibata, N.; Ohmoto, K.; Fujiwara, Toshiyoshi; Yamamoto, S.; Tanaka, N.

In: Cell Transplantation, Vol. 11, No. 5, 2002, p. 421-428.

Research output: Contribution to journalArticle

Kunieda, T, Kobayashi, N, Sakaguchi, M, Okitsu, T, Totsugawa, T, Watanabe, T, Matsumura, T, Maruyama, M, Noguchi, H, Takesue, M, Shibata, N, Ohmoto, K, Fujiwara, T, Yamamoto, S & Tanaka, N 2002, 'Transduction of immortalized human hepatocytes with p21 to enhance differentiated phenotypes', Cell Transplantation, vol. 11, no. 5, pp. 421-428.
Kunieda T, Kobayashi N, Sakaguchi M, Okitsu T, Totsugawa T, Watanabe T et al. Transduction of immortalized human hepatocytes with p21 to enhance differentiated phenotypes. Cell Transplantation. 2002;11(5):421-428.
Kunieda, T. ; Kobayashi, N. ; Sakaguchi, Masakiyo ; Okitsu, T. ; Totsugawa, T. ; Watanabe, T. ; Matsumura, T. ; Maruyama, M. ; Noguchi, H. ; Takesue, M. ; Shibata, N. ; Ohmoto, K. ; Fujiwara, Toshiyoshi ; Yamamoto, S. ; Tanaka, N. / Transduction of immortalized human hepatocytes with p21 to enhance differentiated phenotypes. In: Cell Transplantation. 2002 ; Vol. 11, No. 5. pp. 421-428.
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AU - Sakaguchi, Masakiyo

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