title: Vitamin D(3)  cryosensitization increases prostate cancer susceptibility to cryoablation via mitochondrial-mediated apoptosis and necrosis.
abstract: OBJECTIVE
  To investigate the effect and molecular mechanisms of action of Vitamin D(3) (VD(3) ) as a neo-adjunctive agent before cryosurgery in an effort to increase treatment efficacy for prostate cancer (CaP). To eliminate the potential for disease recurrence that exists at the periphery of the freeze lesion, where temperatures may be insufficient to destroy both androgen-sensitive (AS) and androgen-insensitive (AI) CaP.
  
  
  METHODS
  Human CaP cells, LNCaP, were each genetically altered to express the AS and AI phenotypes and subjected to VD(3) treatment and freezing in an in vitro and tissue-engineered model. Cell viability, caspase inhibitor and western blot studies were used to determine the basis of the different responses of AI and AS cells to VD(3) cryosensitization.
  
  
  RESULTS
  VD(3) was found to be a highly effective cryosensitizer, resulting in a >50% overall increase in cell death after -15 °C freezing. Fluorescence microscopy, western blot analysis and caspase protease assays confirmed that the increased activation of apoptosis was modulated through a mitochondrial-mediated pathway. Caspase inhibition studies showed that apoptosis played an integral role in cell death, with VD(3) cryosensitivation-induced apoptotic events responsible for >30% of the overall cell death after -15 °C freezing.
  
  
  CONCLUSIONS
  The present study suggests that the use of VD(3) as a cryosensitizer increases cryoablation efficacy through the increased activity of apoptosis as well as through necrosis. The data show that through VD(3) treatment the overall level of AI CaP cell tolerance to freezing is reduced to a level similar to that of AS CaP. VD(3) pre-treatment in conjunction with cryoablation may increase treatment efficacy and reduce disease recurrence for CaP patients.
authors: Department of Biological Sciences, Binghamton University, Binghamton, NY, USA. jmbaust@cellpreservation.com
authors: John M
authors: JM
authors: Baust
authors: Daniel P
authors: DP
authors: Klossner
authors: Anthony
authors: A
authors: Robilotto
authors: Robert G
authors: RG
authors: Vanbuskirk
authors: Andrew A
authors: AA
authors: Gage
authors: Vladimir
authors: V
authors: Mouraviev
authors: Thomas J
authors: TJ
authors: Polascik
authors: John G
authors: JG
authors: Baust
mesh_terms: D017209:Apoptosis / Q000187:drug effects; D001681:Biological Assay; D015153:Blotting, Western; D020169:Caspases / Q000378:metabolism; D016923:Cell Death / Q000187:drug effects; D002762:Cholecalciferol / Q000494:pharmacology* / Q000627:therapeutic use; D003452:Cryosurgery / Q000379:methods*; D006801:Humans; D008297:Male; D008856:Microscopy, Fluorescence; D008928:Mitochondria / Q000187:drug effects; D011471:Prostatic Neoplasms / Q000601:surgery*; D016896:Treatment Outcome; D014407:Tumor Cells, Cultured; D014815:Vitamins / Q000494:pharmacology* / Q000627:therapeutic use
chemical_list: D014815:Vitamins; D002762:Cholecalciferol; D020169:Caspases