The Penetratin Sequence in the Anticancer PNC-28 Peptide Causes Tumor Cell Necrosis Rather Than Apoptosis of Human Pancreatic Cancer Cells

详细信息    查看全文

• 作者：Wilbur B. Bowne MD (1) (2)
  Kelley A. Sookraj MD (1) (2)
  Michael Vishnevetsky (3)
  Victor Adler MD ; PhD (4)
  Ehsan Sarafraz-Yazdi MS (5)
  Sunming Lou PhD (5)
  Jesco Koenke MS (6)
  Vadim Shteyler (3)
  Kamran Ikram MD (1) (2)
  Michael Harding (4)
  Martin H. Bluth MD ; PhD (7)
  Mou Ng PhD (3)
  Paul W. Brandt-Rauf MD (8)
  Raqibul Hannan PhD (3)
  Stephan Bradu MD ; PhD (9)
  Michael E. Zenilman MD (1)
  Josef Michl MD (10)
  Matthew R. Pincus MD ; PhD (3) (4)
  
• 刊名：Annals of Surgical Oncology
• 出版年：2008
• 出版时间：December 2008
• 年：2008
• 卷：15
• 期：12
• 页码：3588-3600
• 全文大小：538KB
• 参考文献：1. Kanovsky M, Raffo A, Drew L et al. Peptides from the amino terminal mdm-2 binding domain of p53, designed from conformational analysis, are selectively cytotoxic to transformed cells. / Proc Natl Acad Sci USA 2001; 98:12438-3 CrossRef
  2. Do TN, Rosal RV, Drew L et al. Preferential induction of necrosis in human breast cancer cells by a p53 peptide derived from the mdm-2 binding site. / Oncogene 2003; 22:1431-4 CrossRef
  3. Pincus MR, Michl J, Bowne W et al. Anti-Cancer Peptides from the ras-p21 and p53 Proteins. / Research Advances in Cancer. Mohan RM (ed). Global Research Network, Kerala, India, pp 65-0, 2007
  4. Rosal R, Pincus MR, Brandt-Rauf PW et al. NMR solution structure of a peptide from the mdm-2 binding domain of the p53 protein that is selectively cytotoxic to cancer cells. / Biochemistry 2004; 43:1754-61 CrossRef
  5. Rosal R, Brandt-Rauf PW, Pincus MR et al. The role of alpha-helical structure in p53 peptides as a determinant for their mechanism of cell death: necrosis versus apoptosis. / Adv Drug Devel Rev 2004; 57:653-0 CrossRef
  6. Michl J, Scharf B, Schmidt A et al. PNC-28, a p53 peptide that is cytotoxic to cancer cells, blocks pancreatic cancer cell growth in?vivo. / Int J Cancer 2006; 119:1577-5 CrossRef
  7. Picksley SM, Spicer JF, Barnes DM et al. The p53-MDM2 interaction in a cancer-prone family, and the identification of a novel therapeutic target. / Acta Oncol 1996; 35:429-4 CrossRef
  8. Bottger V, Bottger A, Howard SF et al. Identification of novel mdm2 binding peptides by phage display. / Oncogene 1996; 13:2141-
  9. Wasylyk C, Salvi R, Argentini M et al. p53 mediated death of cells overexpressing MDM2 by an inhibitor of MDM2 interaction with p53. / Oncogene 1999; 18:1921-4 CrossRef
  10. Chene P, Fuchs J, Carena I et al. Study of the cytotoxic effect of a peptidic inhibitor of the p53-hdm2 interaction in tumor cells. / FEBS Lett 2002; 529:293- CrossRef
  11. Garcia-Echeverria C, Furet P, Chene P. Coupling of the antennapedia third helix to a potent antagonist of the p53/hdm2 protein-protein interaction. / Bioorg Med Chem Lett 2001; 11:2161- CrossRef
  12. Harbour JW, Worley L, Ma D et al. Transducible peptide therapy for uveal melanoma and retinoblastoma. / Arch Ophthalmol 2002; 120:1341-
  13. Sambrook J, Russell DW. / Molecular Cloning: A Laboratory Manual. Cold Spring Harbor, NY: CSHL, 2001
  14. Kanovsky M, Michl J, Botzolaki G et al. Peptides, designed from molecular modeling studies of the ras-p21 protein, induce phenotypic reversion of a pancreatic carcinoma cell line but have no effect on normal pancreatic acinar cell growth. / Cancer Chemother Pharmacol 2003; 52:202- CrossRef
  15. van den Eijnde SM, Boshart L, Baehrecke EH et al. Cell surface exposure of phosphatidylserine during apoptosis is phylogenetically conserved. / Apoptosis 1998; 3:9-6 CrossRef
  16. Camus S, Higgins M, Lane D et al. Differences in the ubiquitination of p53 by mdm2 and the HPV protein E6. / FEBS Lett 2003; 536:220- CrossRef
  
• 作者单位：Wilbur B. Bowne MD (1) (2)
  Kelley A. Sookraj MD (1) (2)
  Michael Vishnevetsky (3)
  Victor Adler MD, PhD (4)
  Ehsan Sarafraz-Yazdi MS (5)
  Sunming Lou PhD (5)
  Jesco Koenke MS (6)
  Vadim Shteyler (3)
  Kamran Ikram MD (1) (2)
  Michael Harding (4)
  Martin H. Bluth MD, PhD (7)
  Mou Ng PhD (3)
  Paul W. Brandt-Rauf MD (8)
  Raqibul Hannan PhD (3)
  Stephan Bradu MD, PhD (9)
  Michael E. Zenilman MD (1)
  Josef Michl MD (10)
  Matthew R. Pincus MD, PhD (3) (4)
  
  1. Department of Surgery, SUNY Downstate Medical Center, 450 Clarkson Avenue, Brooklyn, NY, 11203, USA
  2. Department of Surgery, New York Harbor VA Medical Center, 800 Poly Place, Brooklyn, NY, 11209, USA
  3. Department of Pathology, SUNY Downstate Medical Center, 450 Clarkson Avenue, Brooklyn, NY, 11203, USA
  4. Department of Pathology and Laboratory Medicine, New York Harbor Veterans Administration Medical Center, 800 Poly Place, Brooklyn, New York, 11209, USA
  5. Molecular Biology Graduate Program, SUNY Downstate Medical Center, 450 Clarkson Avenue, Brooklyn, NY, 11203, USA
  6. Department of Biology, Columbia University, New York, NY, USA
  7. Department of Pathology, Wayne State Medical Center, Detroit, MI, USA
  8. Department of Environmental Sciences, Columbia College of Physicians and Surgeons, 60 Haven Avenue, New York, NY, 10032, USA
  9. Department of Dermatology, SUNY Downstate Medical Center, 450 Clarkson Avenue, Brooklyn, NY, 11203, USA
  10. Departments of Pathology, Microbiology, Anatomy, and Cell Biology, SUNY Downstate Medical Center, 450 Clarkson Avenue, Brooklyn, NY, 11203, USA
  

文摘

Background PNC-27 and PNC-28 are p53-derived peptides from the human double minute (hdm-2) binding domain attached to penetratin. These peptides induce tumor cell necrosis of cancer cells, but not normal cells. The anticancer activity and mechanism of PNC-28 (p53 aa17-6-penetratin) was specifically studied against human pancreatic cancer. Methods MiaPaCa-2 cells were treated with PNC-28. Necrosis was determined by measuring lactate dehydrogenase (LDH) and apoptosis as assayed for measuring elevation of proapoptotic proteins. PNC-29, an unrelated peptide, and hdm-2-binding domain p53 aa12-26 without penetratin (PNC-26) were used as controls. Since there is evidence that penetratin is required for tumor cell necrosis, we tested “naked-p53 peptide without penetratin by transfecting a plasmid that encodes p53 aa17-6 segment of PNC-28 into MiaPaCa-2 and an untransformed rat pancreatic acinar cell line, BMRPA1. Time-lapse electron microscopy was employed to further elucidate anticancer mechanism. Results Treatment with PNC-28 does not result in the elevation of proapoptotic proteins found in p53-induced apoptosis, but elicits rapid release of LDH, indicative of tumor cell necrosis. Accordingly, we observed membrane pore formation and dose-dependent killing. In direct contrast, transfected MiaPaCa-2 cells underwent apoptosis, and not necrosis, as evidenced by expression of high levels of caspases-3 and 7 and annexin V with background levels of LDH. Conclusion These results suggest that PNC-28 may be effective in treating human pancreatic cancer. The penetratin sequence appears to be responsible for the fundamental change in the mechanism of action, inducing rapid necrosis initiated by membrane pore formation. Cancer cell death by apoptosis was observed in the absence of penetratin.