姜黄素对雄激素非依赖性前列腺癌细胞Notch1表达和分布的影响

目的探讨姜黄素对雄激素非依赖性前列腺癌PC3细胞增殖、凋亡及Notch1表达水平、分布定位的影响。方法体外培养PC3细胞给予不同浓度的姜黄素处理，以MTT法检测细胞增殖率；以RT—PCR检测细胞Notch1 mRNA表达情况；流式细胞术检测细胞凋亡率；透射电镜观察细胞超微结构的变化；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。结果姜黄素抑制PC3细胞的增殖呈时间和剂量依赖性（P〈0．01）；RT—PCR、免疫荧光细胞化学实验结果表明PC3细胞株有Notch1 mRNA和蛋白的表达，且随姜黄素浓度升高而增高（P〈0．05）；FCM显示PC3细胞凋亡率随姜黄素浓度升高而增加（P〈0．01）；电镜观察到典型凋亡形态学变化；激光共聚焦检测结果显示Notch1不仅有胞膜、胞质定位，胞核也有少量表达，随姜黄素浓度升高胞核内表达增加。结论姜黄素抑制PC3细胞增殖并诱导凋亡，呈时间和剂量依赖性。PC3细胞对姜黄素的反应性可能与Notch1的表达和核易位激活有关。

Effect of Curcumin on the Expression and Distribution of Notch1 in Androgen Independent Prostate Cancer Cell Line PC3 in vitro

Objective To investigate the effects of curcumin on the proliferation and apoptosis and the expression and distribution of Notch1 in androgen independent prostate cancer cell line(PC3).Methods After in vitro cultured PC3 cells were induced by different concentrations of curcumin respectively,MTT assay was applied to determine the proliferation at different time points;Notch1 mRNA expression was detected by RT-PCR;Apoptosis rate was assayed by flow cytometry;The morphological changes were observed under the inverted electron microscopy;Laser scanning confocal microscope immunofluorescence analysis was performed to detect the expression and the sub-cellular localization of Notch1 protein.Results The growth of PC3 cells was suppressed obviously by curcumin in a dose-and time-dependent manner(P<0.01).RT-PCR and fluorescent immunocytochemistry assay revealed that the expression of Notch1 mRNA and protein was detected in PC3 cells and increased with the increase of curcumin concentration(P<0.05).The results of confocal microscopic immunofluorescence assay showed that the Notch1 was not only localized in membrane and cytoplasm but also a little in nucleolus of PC3 cells.The quantity of Notch1 in the nuclei was gradually increased in a dose-dependent manner of curcumin.Conclusion Curcumin could suppress the growth of PC3 cells and promote the apoptosis in a dose-and time-dependent manner.Notch1 was located mainly in membrane and cytoplasm of PC3 cells.The expression of Notch1 and nuclear translocation was gradually increased with the increase of curcumin.The reactivity of the PC3 cells to curcumin may be due to the intracellular location of Notch1 receptors and specific signaling pathway.