伊曲康唑对前列腺癌PC-3细胞凋亡的影响及其机制  

Effects and mechanism ofitraconazole on prostate cancer PC-3 cell apoptosis

作　　者：赵中伟[1] 羊丽丽[1] 纪建松[1] 郑丽云[1] 方世记[1] 王均炉[2] 

机构地区：[1]温州医科大学附属第五医院、丽水市中心医院介入诊疗中心,丽水323000 [2]温州医科大学附属第一医院麻醉科,丽水323000

出　　处：《中华医学杂志》2016年第39期3160-3163,共4页National Medical Journal of China

基　　金：国家自然科学基金（81573657）;浙江省科技厅公益性技术应用研究计划项目（2014c3147）

摘　　要：目的 探讨伊曲康唑对前列腺癌PC-3细胞增殖的影响及其机制.方法 将前列腺癌PC-3细胞分为对照组、伊曲康唑组、伊曲康唑＋神经酰胺合成酶1（CerS1）-shRNA组（伊曲康唑＋CerS1-shRNA组）和伊曲康唑＋1-苯基-2-癸酰基氨基-3-吗啉代-1-丙醇（PDMP）组（伊曲康唑＋PDMP组）,运用噻唑蓝染色法检测PC-3细胞生长,Annexin V-FITC/PI双染法检测细胞凋亡,高效液相色谱法检测细胞内神经酰胺含量,蛋白免疫印迹方法检测Bax、Bcl-2、cleaved-caspase3和Akt、mTORC1及其磷酸化的表达水平.结果 终浓度为0、5、10、20μmol/L伊曲康唑诱导细胞凋亡和神经酰胺含量增高,各浓度细胞凋亡率分别为3.23％±1.32％、5.87％ ±2.45％、23.22％±5.29％、48.57％±8.37％;其中10和20 μmol/L时与对照组比较,差异有统计学意义（均P＜0.05）.神经酰胺含量百分率分别为100％、109％±18％、156％±12％、197％ ±22％;其中10和20 μmol/L时与对照组比较,差异均有统计学意义（均P ＜0.05）.蛋白免疫印迹结果显示伊曲康唑组和伊曲康唑＋PDMP组Bax、cleaved-caspase 3的表达明显高于对照组,而Bcl-2表达明显少于对照组;伊曲康唑＋CerS1-shRNA组Bax、cleaved-caspase 3表达明显低于伊曲康唑组,而Bcl-2表达明显高于伊曲康唑组.5、10、20 μmol/L伊曲康唑处理的PC-3细胞p-Akt、p-mTORC1的表达明显低于对照组;伊曲康唑＋CerS-1-shRNA组p-Akt、p-mTORC1的表达明显高于伊曲康唑组.结论 伊曲康唑通过增加细胞内神经酰胺含量抑制PC-3细胞生长和诱导细胞凋亡,可能与神经酰胺增加caspase 3活化、Bax及降低Bcl-2表达和抑制Akt-mTORC信号通路有关.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（all P 〈 0.05）.Western blot analysis showed that the Bax,cleavedcaspase 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.

关 键 词：伊曲康唑 前列腺肿瘤 神经酰胺类 bcl-X蛋白 

分 类 号：R737.25[医药卫生—肿瘤]