自噬在熊果酸诱导前列腺癌PC3细胞凋亡中的作用机制研究

The Mechanism of Autophagy in Ursolic Acid Inducing Prostatic Cancer PC3 Cells Apoptosis

为了探讨自噬在熊果酸抑制前列腺癌PC3细胞凋亡中的作用机制,PC3细胞培养至对数生长期后,以无糖、无氨基酸培养液代替原培养液培养细胞,并用不同浓度的熊果酸进行干预。72 h后收集细胞,采用透射电镜和免疫荧光技术观察PC3细胞自噬情况;Western Blot检测ATG5和Beclin-1蛋白表达;Elisa法测定Caspase-3、Caspase-8和Caspase-9含量;流式细胞技术检测PC3细胞凋亡情况。结果表明,PC3细胞饥饿72 h后,细胞自噬明显增强。与对照组比较,熊果酸作用后的细胞自噬程度明显减弱;ATG5和Beclin-1蛋白表达显著减少(P<0.05);Caspase-3、Caspase-8和Caspase-9含量显著增加(P<0.05);PC3细胞凋亡率极显著升高(P<0.01)。实验初步揭示,熊果酸可通过抑制饥饿状态的前列腺癌PC3细胞自噬,并进一步通过促进凋亡因子的分泌诱导其凋亡。

This paper is main to explore the mechanism of autophagy in ursolic acid inducing prostatic cancer PC3 cells apoptosis. PC3 cells were cultured until logarithmic growth phase,then culture medium was insteaded by sugar and amino acid free medium,and ursolic acid of different concentration was joined to intervene. PC3 cells were collected after 72 h,transmission electron microscope and immunofluorescence were used to detect PC3 cells autophagy status; expression of ATG5 and Beclin-1 proteins were detected by western blot; Caspase-3、Caspase-8 and Caspase-9 were detected by Elisa method; and the apoptosis was detected by flow cytometry. The results showed that the autophagy of PC3 cells aggravated obviously after 72 hours. Compared with the control group,cells autophagy in ursolic acid group weaked obviously; The expression of ATG5 and Beclin-1 proteins were reduced markedly（ P 〈 0. 05）,and the content of Caspase-3、Caspase-8 and Caspase-9 were reduced markedly as well（ P 〈 0. 05）. What’s more,the apoptosis rate rised extremely significant（ P〈 0. 05）. Our datas indicated that Ursolic acid may inhibit autophagy of hungry PC3 cells,which might induce cells apoptosis via facilitating the secretion of apoptosis factors.