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2008 Annual Meeting Abstracts
Identification Of Novel Mechanisms Involved In Pro-apoptotic, Neuroprotective, And Anti-metastatic Effects Of Cryo-shock Conditioned Media On Prostate Cancer Cells
Mini Varghese, Seema Gupta*, Mohammed M Shareef*, Daniel B Rukstalis, Mansoor M Ahmed*
Geisinger Health System, Danville, PA
Introduction: Cryotherapy is an accepted modality for prostate cancer treatment. In this study, we designed experiments to identify factors induced in response to cryo-shock temperatures that may have potential direct and bystander effects.
Methods: PC-3 cell cultures were subjected to temperatures of 4°C, 0°C, -5°C, -10°C, and -20°C. Cryo Shock Conditioned Media (CSCM) was collected after treatment and underwent (1) protein precipitation and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) coupled with Matrix Assisted Laser Desorption/Ionization-Time Of Flight (MALDI-TOF) sequencing; and (2) TransSignal Human Cytokine Antibody Array analysis. RNA was isolated for Real-Time Polymerase Chain Reaction Apoptotic SuperArray gene analysis. The bystander effect of CSCM was assessed in growing PC-3 cells using Real-Time Cell Electronic Sensing system.
Results: SDS-PAGE analysis revealed a unique band at the 18 kilodalton range in CSCM from -20°C cells. MALDI-TOF sequencing of this band revealed the presence of CypA (cyclophilin A) and NM23 (nonmetastatic protein 23). Significant induction of interleukin-1α (IL-1α) was shown in Cytokine Antibody Array analysis. Apoptotic SuperArray gene analysis revealed significant up-regulation of NOD1. 80% reduction in cell growth was observed in PC-3 cells exposed to CSCM from -10°C cells.
Conclusions: CypA has been shown to facilitate neuroprotective mechanisms. IL-1α has been reported in animal models to cause tumor regression. NM23 may suppress metastatic deposits. NOD-1 has been linked to caspase induced cell death. This study reveals the presence of multiple factors in CSCM that can contribute to effective prostate tumor management through apoptosis, neuronal protection, and suppression of metastasis.
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