论文部分内容阅读
目的探讨胡桃醌对宫颈癌He La细胞促凋亡作用的机制,及其相关的信号转导通路。方法选取处于对数生长期的He La细胞将其分为空白对照组和30μmol/L胡桃醌组。采用MTT法观察胡桃醌对不同时间点He La细胞的抑制作用;Hoechst 33258试剂盒,流式细胞仪测定胡桃醌对不同时间点He La细胞凋亡的影响;Western blotting检测凋亡相关蛋白Bcl-2、Bax和Caspase-3,磷脂酰肌醇3-激酶(PI3K)/Akt通路中AKt及p Akt的表达。结果MTT实验显示,与对照组比较,各时间点He La细胞具有抑制作用,且差异显著均有统计学意义(P<0.05,P<0.01);Hoechst 33258检测表明,30μmol/L胡桃醌处理细胞4,8,12 h后可出现明显的细胞核典型凋亡细胞形态;细胞凋亡检测表明,30μmol/L胡桃醌培养不同时间后,与对照组相比早期凋亡率明显增高;Western blotting检测显示,与正常对照组相比,30μmol/L胡桃醌处理细胞12h后,He La细胞Bcl-2,p Akt蛋白的表达明显降低,而Bax、Cleave-Caspase-3,Akt蛋白表达明显升高。结论胡桃醌通过抑制PI3K/Akt通路,进而促进He La细胞凋亡。
Objective To investigate the mechanism of juglone on promoting apoptosis of cervical cancer HeLa cells and its related signal transduction pathway. Methods Hela cells in logarithmic growth phase were divided into blank control group and 30μmol / L juglone group. MTT assay was used to observe the inhibitory effect of juglone on HeLa cells at different time points; Hoechst 33258 kit was used to determine the effect of juglone on apoptosis of HeLa cells at different time points by flow cytometry; Western blotting was used to detect the expression of Bcl- 2, Bax and Caspase-3, phosphoinositide 3-kinase (PI3K) / Akt pathway. Results MTT assay showed that compared with the control group, He La cells had inhibitory effect at all time points, and the difference was statistically significant (P <0.05, P <0.01). Hoechst 33258 showed that 30μmol / L juglone treated cells After 4, 8, and 12 h, obvious apoptotic cell morphologies were observed. Apoptosis assay showed that the early apoptotic rate of 30 μmol / L juglone was significantly higher than that of the control group at different time points. The results of Western blotting Compared with the normal control group, the expression of Bcl-2, p Akt and Bax, Cleave-Caspase-3 and Akt proteins were significantly increased in Hela cells treated with 30μmol / L juglone for 12h. Conclusion Juglone can promote the apoptosis of HeLa cells by inhibiting the PI3K / Akt pathway.