摘要
目的 观察丙戊酸(VPA)诱导前列腺癌PC3细胞发生自噬性死亡的现象,初步探讨其发生的可能机制.方法 VPA作用前列腺癌PC3细胞后,应用CCK-8试剂盒检测细胞生存率,透射电子显微镜及细胞免疫荧光测定仪观察细胞超微结构及自噬水平,Westem-blotting检测PC3细胞内自噬特异性蛋白-Ⅱ(LC3-Ⅱ)及p-Akt蛋白的表达水平.结果 经VPA处理后,前列腺癌PC3细胞活性短期内(≤4d)未受到明显抑制.透射电子显微镜观察可见,前列腺癌PC3细胞质内有大量自噬体和自噬溶酶体,其自噬水平随VPA作用时间的延长逐渐增强.Western-blotting检测表明,前列腺癌PC3细胞LC3-Ⅱ表达水平随VPA作用时间的延长明显升高,而p-Akt表达水平则明显降低.结论 VPA可诱导前列腺癌PC3细胞发生自噬,其诱导前列腺癌PC3细胞发生自噬的机制可能与阻断Akt/mTOR信号转导通路有关.
Abstract
Objective To observe autophagy of prostate cancer PC3 cells induced by valproic acid (VPA)and explore the possible mechanism. Methods The prostate cancer PC3 cell line treated with VPA was used in this study. Cell survival rate was determined by cell counting kit-8. Autophagosomes were observed under a transmission electronic microscope. The intensity of autophagy was analyzed by immunofluorescence. Expression levels of LC3-Ⅱ and p-Akt were measured by Western blot after VPA treatment. Results Viability of the prostate cancer PC3 cell line treated with acute VPA was not significantly inhibited in a short time( ≤4 d). Autophagosomes and autolysosomes were observed in the VPA-treated prostate cancer PC3 cell line under a transmission electronic microscope. Intensity of autophagy was in a time-dependent manner. Expression level of the related protein LC3- Ⅱ was elevated, while expression level of p-Akt declined following VPA treatment in the prostate cancer PC3 cell line. Conclusions VPA may induce autophagy in the prostate cancer PC3 cell line. The mechanism may be related to the disruption of the Akt/mTOR signaling pathway.
NETL
NSTL
维普
万方数据