title: [Effects and mechanism ofitraconazole on prostate cancer PC-3 cell apoptosis].
abstract: Objective:To investigate the effects and mechanisms of itraconazole on prostate cancer PC-3 cells proliferation.Methods:The PC-3 cells were divided into four group: control group, itraconazole group, itraconazole+ CerS-1-shRNA group and itraconazole+ PDMP group.3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2-H-tetrazolium bromide(MTT)assay was used to detect the growth of PC-3 cells.Apoptosis was detected by Annexin V-FITC /PI.The intracellular ceramide production was assayed by high performance liquid chromatography(HPLC). The expression of Bax, Bcl-2, cleaved-caspase3 and the expression and phosphorylation of Akt, mTORC1 were detected by Western blot.Results:After treatment with 0, 5, 10, 20 μmol/L itraconazole, apoptosis rate was 3.23%±1.32%, 5.87%±2.45%, 23.22%±5.29%, 48.57%±8.37%.The percentage content of ceramide was 100%, 109%±18%, 156%±12%, 197%±22%.Compared with the control group, there were statistically differences when the concentration of itraconazole were 10 and 20 μmol/L(allP<0.05). Western blot analysis showed that the Bax, cleaved-caspase 3 expression of itraconazole group and itraconazole+ PDMP group was significantly higher than control group, while Bcl-2 expression was significantly lower than the control; the Bax, cleaved-caspase 3 expression ofitraconazole+ CerS1-shRNA group was significantly lower than itraconazole group, while Bcl-2 expression was significantly higher than the itraconazole group.After 5, 10, 20 μmol/L itraconazole treatment, the expression of p-Akt and p-mTORC1 were significantly lower than the control group; the expression of p-Akt and p-mTORC1 in itraconazole+ CerS-1-shRNA group were significantly higher than itraconazole group.Conclusion:Itraconazole induces apoptosis of PC-3 cell through increasing the intracellular ceramide content, which might relate to upregulation of cleavage-caspase 3 and Bax, downregulation of Bcl-2 and inactivation of Akt-mTORC signal pathway.
authors.affiliation: Department of Interventional Radiology, the Fifth Affiliated Hospital of Wenzhou Medical University, Lishui 323000, China.
authors.forename: Z W
authors.initials: ZW
authors.lastname: Zhao
authors.forename: L L
authors.initials: LL
authors.lastname: Yang
authors.forename: J S
authors.initials: JS
authors.lastname: Ji
authors.forename: L Y
authors.initials: LY
authors.lastname: Zheng
authors.forename: S J
authors.initials: SJ
authors.lastname: Fang
authors.forename: J L
authors.initials: JL
authors.lastname: Wang
mesh_terms: D017209:Apoptosis; D053148:Caspase 3; D020169:Caspases; D045744:Cell Line, Tumor; D006801:Humans; D008297:Male; D000076222:Mechanistic Target of Rapamycin Complex 1; D010766:Phosphorylation; D011471:Prostatic Neoplasms; D015398:Signal Transduction
chemical_list: D000076222:Mechanistic Target of Rapamycin Complex 1; C505407:CASP3 protein, human; D053148:Caspase 3; D020169:Caspases