摘要
目的探讨桑根酮C诱导雄激素非依赖型前列腺癌PC3细胞凋亡的作用及其可能机制。方法 MTT法检测桑根酮C对PC3细胞増殖的抑制作用:实验分6组,分别加入不同浓度的桑根酮C(0、1、5、20、50、100μmol/L),作用24 h检测细胞生长抑制率;20μmol/L桑根酮C加入PC3细胞,分别于0、6、12、24、48 h收集细胞,MTT法检测生长抑制率。流式细胞技术检测细胞凋亡率:20μmol/L桑根酮C加入PC3细胞,分别于0、12、24、48 h收集细胞检测凋亡率。荧光分光光度计检测caspase 3的活性:20μmol/L桑根酮C作用PC3细胞,分别在0、6、12、18、24、48 h收集细胞,检测caspases 3的活性。MTT法检测caspase 3、caspase 8及caspase 9抑制剂对桑根酮C抑制PC3细胞增殖的影响:实验分4组,分别加入DMSO、z-DEVD-fmk、z-LEHD-fmk及z-IETD-fmk,1 h加入桑根酮C,24 h收集细胞,MTT法计算细胞生长抑制率。结果桑根酮C能抑制PC3细胞的增殖,呈剂量、时间依赖性,桑根酮C 1、5、20、50、100μmol/L作用PC3细胞24 h的增值抑制率分别为:4.86%、12.92%、52.34%、82.05%、87.45%,1μmol/L组与对照组相比差异无统计学意义(P>0.05),其他组与对照组相比差异有统计学意义(P<0.05)。半数有效抑制浓度IC50为18.76μmol/L。20μmol/L桑根酮C作用PC3细胞,分别在6、12、24、48、72 h检测增值抑制率为:10.57%、27.09%、51.88%、80.73%、87.99%,与对照组相比差异均有统计学意义(P<0.05)。20μmol/L桑根酮C可诱导PC3细胞的凋亡,在12、24、48 h的凋亡率分别为:7.43%、20.91%、37.56%,与对照组相比差异有统计学意义(P<0.05)。检测caspases 3的活性18 h比对照组增加了7.6倍,应用caspase抑制剂后发现caspase 3及caspase 9的抑制剂可显著逆转桑根酮C对PC3细胞增殖的抑制作用,抑制率从52.48%降到24.29%及28.81%,与对照组相比差异有统计学意义(P<0.01)。而caspase8抑制剂作用不明显,与对照组相比差异没有统计学意义(P>0.05)。结论桑根酮C可通过激活caspase 3及caspase 9途径诱导PC3细胞的凋亡。
Objective To investigate the effects of Sanggenon C in inducing apoptosis of prostate cancer PC3 cell line and explore the underlying mechanism.Methods The proliferation of PC3 cells treated for 24 h with 1,5,20,50,and 100 μmol/L sanggenon C or treated with 20 μmol/L Sanggenon C for 0,6,12,24 and 48 h was evaluated using MTT assay.Flow cytometry was performed for analysis of apoptosis of PC3 cells after exposure to sanggenon C with different treatment protocols,and the activity of caspase 3 was detected using spectrofluorometry.The inhibitory effect of sanggenon C on PC3 cells pretreated with DMSO,z-DEVD-fmk,z-LEHD-fmk or z-IETD-fmk for 1 h was detected by MTT assay.Results Sanggenon C inhibited the proliferation of PC3 cells in a dose-and time-dependent manner (P〈0.05 except for 1 μmol/L group) with a 24-h IC50of18.76 μmol/L.Sanggenon C at 20 μmol/L caused inhibition rates of PC3 cells of 10.57%,27.09%,51.88%,80.73% and 87.99%after treatment for 6,12,24,48,and 72 h,respectively (P〈0.05),and resulted in apoptosis rates of 7.43%,20.91% and 37.56% at12 h,24 h and 48 h,respectively.Sanggenon C significantly increased caspase-3 activity in the cells,and its effect on PC3 cell proliferation was partially reversed by caspase 3 and caspase 9 inhibitors.Conclusion Sanggenon C can dose-dependently induce growth inhibition and apoptosis of PC3 cells possibly by activating caspase 9 and caspase 3 pathways.
作者
周萍
董晓先
汤平
ZHOU Ping DONG Xiaoxian TANG Ping(Basic Medical Research Center Department of Pathophysiology, Guangzhou Medical University, Guangzhou 511436, China Department of Urology, Guangzhou First People's Hospital, Guangzhou 510180, China)
出处
《南方医科大学学报》
CAS
CSCD
北大核心
2017年第9期1206-1210,共5页
Journal of Southern Medical University
基金
国家自然科学基金(81072091/H1619)~~