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Anti-Cancer Peptide PNC-27 Kills Cancer Cells by Unique Interactions with Plasma Membrane-Bound hdm-2 and with Mitochondrial Membranes Causing Mitochondrial Disruption

Study · human · Annals of clinical and laboratory science · 2024 · PMID 38802154

Plain-language summary

Paraphrased from the published abstract below — not a verdict on whether anything works.

This in vitro study used human MIA-PaCa-2 pancreatic carcinoma cells to examine how the peptide PNC-27 interacts with membrane-bound HDM-2 and mitochondria. Cells were incubated with PNC-27 alongside a monoclonal antibody targeting the p53 binding site (residues 1-109) of HDM-2, and with mitotracker and lysotracker dyes to assess organelle function; immuno-electron microscopy (IEM) with gold-labeled anti-PNC-27 antibody was also performed on mitochondria. The antibody blocked PNC-27-induced cancer cell necrosis, while negative control immune serum did not. Mitochondria of PNC-27-treated cells failed to retain mitotracker dye while lysosomes retained lysotracker dye, and IEM showed gold particles on mitochondrial membranes. The authors report that PNC-27 binds the p53 binding site of HDM-2, inducing transmembrane pore formation, and that the peptide also binds mitochondrial membranes, associated with their disruption.

Abstract

We have previously shown that the anti-cancer peptide PNC-27 kills cancer cells by co-localizing with membrane-expressed HDM-2, resulting in transmembrane pore formation causing extrusion of intracellular contents. We have also observed cancer cell mitochondrial disruption in PNC-27-treated cancer cells. Our objectives are to determine: 1. if PNC-27 binds to the p53 binding site of HDM-2 (residues 1-109) in the cancer cell membrane and 2. if this peptide causes selective disruption of cancer cell mitochondria. For aim 1, we incubated MIA-PaCa-2 human pancreatic carcinoma cells with PNC-27 in the presence of a monoclonal antibody against the amino terminal p53 binding site of HDM-2 to determine if it, but not negative control immune serum, blocks PNC-27-induced tumor cell necrosis. For the second aim, we incubated these cells with PNC-27 in the presence of two specific dyes that highlight normal organelle function: mitotracker for mitochondria and lysotracker for lysosomes. We also performed immuno-electron microscopy (IEM) with gold-labeled anti-PNC-27 antibody on the mitochondria of these cells treated with PNC-27. Monoclonal antibody to the p53 binding site of HDM-2 blocks PNC-27-induced cancer cell necrosis, whereas negative control immune serum does not. The mitochondria of PNC-27-treated cancer cells fail to retain mitotracker dye while their lysosomes retain lysotracker dye. IEM of the mitochondria cancer cells reveals gold particles present on the mitochondrial membranes. PNC-27 binds to the p53 binding site of HDM-2 (residues 1-109) inducing transmembrane pore formation and cancer cell necrosis. Furthermore, this peptide enters cancer cells and binds to the membranes of mitochondria, resulting in their disruption.

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