大黄素通过逆转Notch信号通路抑制雄激素非依赖前列腺癌细胞PC3增殖和诱导凋亡研究  

Emodin inhibits androgen independent prostate cancer cell line PC3 growth in vitro by inversion Notch receptor expression

作　　者：邓刚[1,2] 马立彬[1,2] 居翔 孟琦[1,2] 于志坚[1,2] 

机构地区：[1]南京医科大学附属杭州医院 [2]杭州市第一人民医院,杭州310006

出　　处：《中华中医药杂志》2014年第4期1253-1257,共5页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：浙江省自然科学基金(No.Y2111329);浙江省医药卫生科技计划(No.2011KYB066);浙江省中医药卫生科技计划(No.2011ZB099);杭州市医疗卫生科研项目(No.20110833B05);浙江省医学会临床科研资金项目(No.2012ZYC-A35)~~

摘　　要：目的:研究大黄素(EM)对雄激素非依赖性前列腺癌PC3细胞的增殖抑制和诱导凋亡作用,并探讨EM是否通过Notch信号通路达到抗肿瘤作用。方法:MTT比色法检测10、20、40、60、80μmol/L浓度EM作用于PC3细胞12、24、36、48、72、96h后细胞活性;24h后流式细胞仪测定细胞周期及凋亡的变化,透射电镜观察细胞超微结构变化;RT-PCR和Western Blot检测PC3细胞内Notch1、Jagged1、VEGF和bFGF mRNA和蛋白的表达;激光共聚焦显微镜免疫荧光分析法评价Notch1蛋白的表达水平和亚细胞定位分布。结果:EM能显著抑制PC3细胞的生长,呈剂量与时间依赖性,不同浓度EM组之间及不同时间组之间差异均有统计学意义(P<0.01);PC3细胞出现有EM剂量依赖性G2/M期阻滞(P<0.01),且各浓度组凋亡细胞比例均显著高于空白对照组(P<0.01),差异有统计学意义;EM作用24h后PC3细胞出现凋亡形态学改变;PC3细胞内Notch1 mRNA和蛋白表达均上升,Jagged1、VEGF、bFGF mRNA和蛋白的表达呈剂量依赖性降低;激光共聚焦检测结果显示PC3细胞中Notch1不仅有胞膜、胞浆定位,胞核也有少量表达,随EM浓度升高胞核内表达增加。结论:EM能显著抑制PC3细胞的生长,促进凋亡;并促进其G2/M期阻滞;Notch1的表达和核易位激活引起VEGF、bFGF表达明显降低,可能是EM抑制前列腺癌细胞生长转移的机制之一。Objective： To explore the anticarcinogenic effects of emodin （EM） on the Notch pathway in androgen independent prostate cancer cell line PC3 in vitro. Methods： After PC3 cells lines were induced by 10, 20, 40, 60, 801amol/L EM respectively, the cell activity after 12, 24, 36, 48, 72, 96h was assayed by MTT. Flow cytometry （FCM） and electronic microscopy were adopted to observe cell cycle and morphological changes of PC3 cells at 24 hours. The Notchl, Jaggedl, VEGF and bFGF gene expression were analyzed by semi-quantitative RT-PCR and the protein expression of these in PC3 cell lines were observed by Western Blot at 24 hours. Laser scanning confocal microscope （LSCM） immunofluorescence analysis was performed to detect the level of expression and the sub-cellular localization of Notchl receptor protein. Results： The growth of PC3 cells was suppressed obviously by EM in dose-dependent and time-dependent manners in vitro. The differences between groups of different concentration and different time were statistically significant （P〈0.01）. Furthermore, EM could arrest the cell cycle of PC3 cells at G~/I phase in a dose-dependent manner （P〈0.01）. The ratios of apoptosis were significantly higher than those of controls （P〈0.01）. EM could lead to characteristic morphological changes of apoptosis in PC3 cells after 24 hours. The expression mRNA and protein of Notchl in PC3 cells marked increased meanwhile these of Jaggedl, VEGF and bFGF showed declining all in a dose-dependent manner, the differences were significant as compared with control groups and among the different groups. As far as the subcellular distribution of Notchl was concerned, the results of LSCM assay showed that the Notchl was not only localized in membrane and cytoplasm but also in nucleolus of PC3 cells, the quantity of Notchl in the nuclei gradually increased in a dose- dependent manner of EM groups. Conclusion： EM can suppress the growth of androgen independent prostate cancer cell lines PC3 and promote their apop

关 键 词：大黄素 前列腺癌细胞 PC3细胞 凋亡 NOTCH1 

分 类 号：R737.25[医药卫生—肿瘤]