A recombinant cell-permeable p53 fusion protein is selectively stabilized under hypoxia and inhibits tumor cell growth.

More than 50% of human tumors contain a mutation in p53. Over 90% of tumors are solid tumors. Solid tumors have low oxygenated regions, called hypoxic regions where the tumor cells are more resistant to radio- and chemo-therapy than their well-oxygenated counterparts. In this study, we constructed a cell-permeable p53 fusion protein with selective stability in the hypoxic region. The fusion protein contained the TAT peptide for transduction across membranes, the oxygen-dependent degradation domain of hypoxia-inducible factor-1alpha and wild-type p53. This protein was effectively delivered into tumor cells where it exerted anticancer activity leading to the inhibition of cancer cell growth in vitro and the reduction of tumor weight in vivo. Hence, the fusion protein can be a novel protein drug for antitumor therapies, especially for hypoxic tumor cells.

[1]  Natalie A. Lissy,et al.  Transduction of full-length TAT fusion proteins into mammalian cells: TAT-p27Kip1 induces cell migration , 1998, Nature Medicine.

[2]  S. Schwarze,et al.  In vivo protein transduction: delivery of a biologically active protein into the mouse. , 1999, Science.

[3]  Ronald A. DePinho,et al.  p53: good cop/bad cop. , 2002, Cell.

[4]  Jian Yu,et al.  The transcriptional targets of p53 in apoptosis control. , 2005, Biochemical and biophysical research communications.

[5]  G. G. Stokes.  "J." , 1890, The New Yale Book of Quotations.

[6]  Adrian L. Harris,et al.  Hypoxia — a key regulatory factor in tumour growth , 2002, Nature Reviews Cancer.

[7]  S. Lowe,et al.  Senescence and tumour clearance is triggered by p53 restoration in murine liver carcinomas , 2011, Nature.

[8]  K. Wiman,et al.  Strategies for therapeutic targeting of the p53 pathway in cancer , 2006, Cell Death and Differentiation.

[9]  Soo‐Young Choi,et al.  Intracellular delivery of p53 fused to the basic domain of HIV-1 Tat. , 2004, Molecules and cells.

[10]  W. El-Deiry,et al.  p53 downstream target genes and tumor suppression: a classical view in evolution , 2007, Cell Death and Differentiation.

[11]  Yu Xia Wang,et al.  A novel therapeutic approach to 6-OHDA-induced Parkinson's disease in rats via supplementation of PTD-conjugated tyrosine hydroxylase. , 2006, Biochemical and biophysical research communications.

[12]  A. Harris,et al.  Hypoxia-inducible factor-1 modulates gene expression in solid tumors and influences both angiogenesis and tumor growth. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[13]  Kevin M. Ryan,et al.  DRAM, a p53-Induced Modulator of Autophagy, Is Critical for Apoptosis , 2006, Cell.

[14]  Michael I. Wilson,et al.  Targeting of HIF-α to the von Hippel-Lindau Ubiquitylation Complex by O2-Regulated Prolyl Hydroxylation , 2001, Science.

[15]  K. Engeland,et al.  p53 downregulates expression of the G1/S cell cycle phosphatase Cdc25A , 2007, Oncogene.

[16]  B. Vogelstein,et al.  p53 mutations in human cancers. , 1991, Science.

[17]  V. Rotter,et al.  Transcription regulation by mutant p53 , 2007, Oncogene.

[18]  Masahiro Hiraoka,et al.  Antitumor effect of TAT-oxygen-dependent degradation-caspase-3 fusion protein specifically stabilized and activated in hypoxic tumor cells. , 2002, Cancer research.

[19]  H. Ryan,et al.  HIF-1 alpha is required for solid tumor formation and embryonic vascularization. , 1998, The EMBO journal.

[20]  G. Kong,et al.  Metastasis‐associated protein 1 enhances stability of hypoxia‐inducible factor‐1α protein by recruiting histone deacetylase 1 , 2006, The EMBO journal.

[21]  Janice M. Y. Brown,et al.  The hypoxic cell: a target for selective cancer therapy--eighteenth Bruce F. Cain Memorial Award lecture. , 1999, Cancer research.

[22]  Hiroshi Harada,et al.  Development of a novel fluorescent imaging probe for tumor hypoxia by use of a fusion protein with oxygen-dependent degradation domain of HIF-1&agr; , 2007, SPIE BiOS.

[23]  M. M. Bradford.  A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. , 1976, Analytical biochemistry.

[24]  Yushu Ma,et al.  The transduction of His-TAT-p53 fusion protein into the human osteogenic sarcoma cell line (Saos-2) and its influence on cell cycle arrest and apoptosis , 2008, Molecular Biology Reports.

[25]  P J Slootweg,et al.  p53 overexpression in oral mucosa in relation to smoking , 1999, The Journal of pathology.

[26]  Y. Nakamura,et al.  The role of p53-target genes in human cancer. , 2000, Critical reviews in oncology/hematology.

[27]  K. Takenaga,et al.  Hypoxia-regulated expression of attenuated diphtheria toxin A fused with hypoxia-inducible factor-1alpha oxygen-dependent degradation domain preferentially induces apoptosis of hypoxic cells in solid tumor. , 2005, Cancer research.

[28]  B. Yue,et al.  Transduction of TAT fusion proteins into the human and bovine trabecular meshwork. , 2006, Investigative ophthalmology & visual science.