title: Cryoablation induced the change of TGF-β pathway in CWR-22RV prostate cancer cell line.
abstract: OBJECTIVE
  To assess the efficacy of TGF-β pathway in the CWR-22RV prostate cancer cell line induced by cryoablation.
  
  
  METHODS
  According to the district in the tumor following cryoablation, the CWR-22RV prostate cancer cells were divided into four groups to construct the freezing model of prostate cancer in cell level: Group A, control cells (the uncovered district), Group B, freezing cells (sub-lethal district away from necrosis), Group C, control cells cultured with 1640 and necrosis supernatant and Group D, freezing cells cultured with 1640 and necrosis supernatant (sub-lethal district close to necrosis). Cell apoptosis was observed by flow cytometry analysis 24 h later. Then supernatant in four groups was extracted to test the concentration of TGF-β by ELISA at the time points of 5, 10, 20, 36, and 48 h. At the same time, intracellular TGF-β, Smad2/3, Smad4 of four groups were detected by Western blot at the time point of 10 h.
  
  
  RESULTS
  In aspect of apoptosis, groups B-D have higher apoptosis rate than group A, group D has more apoptosis cells than group B and C. This was verified that the model was successful. Moreover, we found that group C has higher delayed apoptosis rate than group A, and group D has higher early apoptosis rate than other groups (P<0.05); compared with group A, C, D, group B has less TGF-β (P<0.05). Group C secrets more TGF-β than that in group A (P<0.05) and group D secrets more TGF-β than that in group C at the time points of 20, 36, 48 h (P<0.05); Group C and D expressed more Smad2, Smad3 and Smad4 than group A and B at the time point of 10 h after treatment. Meanwhile, cells in group D expressed more Smads than group C.
  
  
  CONCLUSIONS
  Cryoablation could promote TGF-β and its pathway, and the more close to the center of the ice ball, this effect is more apparent.
authors.affiliation: Department of Interventional Therapy, Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center of Cancer, Key Laboratory of Cancer Prevention and Therapy, Tianjin 300060, China.
authors.forename: Tang
authors.initials: T
authors.lastname: Nailing
authors.affiliation: Department of Interventional Therapy, Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center of Cancer, Key Laboratory of Cancer Prevention and Therapy, Tianjin 300060, China. Electronic address: cjr_guozh1@163.com.
authors.forename: Guo
authors.initials: G
authors.lastname: Zhi
authors.affiliation: Department of Interventional Therapy, Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center of Cancer, Key Laboratory of Cancer Prevention and Therapy, Tianjin 300060, China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Xueling
mesh_terms: D017209:Apoptosis / Q000187:drug effects; D045744:Cell Line, Tumor; D003452:Cryosurgery / Q000379:methods*; D006801:Humans; D008297:Male; D011471:Prostatic Neoplasms / Q000378:metabolism* / Q000601:surgery*; D015398:Signal Transduction / Q000187:drug effects; D051899:Smad2 Protein / Q000378:metabolism; D051900:Smad3 Protein / Q000378:metabolism; D051901:Smad4 Protein / Q000378:metabolism; D016212:Transforming Growth Factor beta / Q000378:metabolism*
chemical_list: C497782:SMAD2 protein, human; C497790:SMAD3 protein, human; C497796:SMAD4 protein, human; D051899:Smad2 Protein; D051900:Smad3 Protein; D051901:Smad4 Protein; D016212:Transforming Growth Factor beta
keywords: Cryoablation; Pathway; Prostate cancer; TGF-β