PNC 27

Cancer Protocol PNC 27 - Human Subjects: Induces Tumor Cell Necrosis. PNC 27 is the anti cancer peptide, which is a membrane active peptide that binds to the HDM 2 protein expressed in the cancer cell membranes of solid tissue tumor cells and induces transmembrane pore formation in cancer, but not in normal cells, resulting in tumor cell necrosis that is independent of p53 activity in these cells. We now extend our study to non solid tissue tumor cells, in this case, a primitive, possible stem cell human leukemia cell line K562 that is also p53 homozygously deleted. Our purpose was twofold: to investigate if these cells likewise express HDM 2 in their plasma membranes and to determine if our anti cancer peptide induces tumor cell necrosis in these non solid tissue tumor cells in a manner that depends on the interaction between the peptide and membrane bound HDM 2. The anti cancer activity and mechanism of PNC 27, which carries a p53 aa12 26 leader sequence connected on its carboxyl terminal end to a trans membrane penetrating sequence or membrane residency peptide MRP , was studied against p53 null K562 leukemia cells. Murine leukocytes were used as a non cancer cell control. Necrosis was determined by measuring the lactate dehydrogenase LDH release and apoptosis was determined by the detection of Caspases 3 and 7. Membrane colocalization of PNC 27 with HDM 2 was analysed microscopically using fluorescently labelled antibodies against HDM 2 and PNC 27 peptides.