胡桃醌诱导HeLa细胞凋亡的机制

doi:10.16098/j.issn.0529-1356.2015.05.011

解剖学报 ›› 2015, Vol. 46 ›› Issue (5): 641-645.doi: 10.16098/j.issn.0529-1356.2015.05.011

胡桃醌诱导HeLa细胞凋亡的机制

张巍¹张若文²朱文赫¹ 李妍¹ 罗军¹赵行宇¹ 徐俊杰¹姜艳霞¹吕士杰^1*

1. 吉林医药学院生物化学教研室，吉林吉林 132013; 2. 北华大学基础医学院，吉林吉林 132013

收稿日期:2015-02-13 修回日期:2015-04-23 出版日期:2015-10-06 发布日期:2015-10-06
通讯作者: 吕士杰 E-mail:jlmpcxlc@163.com
基金资助:
聚酮类化合物Talaroconvolutin A的全合成及抗肿瘤活性研究;胡桃醌对宫颈癌细胞株的促凋亡作用机制及与PI3K/AKt通路关系的研究

Effects of juglone on the proliferation and apoptosis of HeLa cells

ZHANG Wei¹ZHANG Ruo-wen²ZHU Wen-he¹ LI Yan¹ LUO Jun¹ ZHAO Xing-yu¹ XU Jun-jie¹ JIANG Yan-xia¹LÜ Shi-Jie ^1*

1. Department of Biochemistry, Jilin Medical College, Jilin Jilin 132013, China; 2. Basic Medical School, Beihua University,Jilin Jilin 132013, China

Received:2015-02-13 Revised:2015-04-23 Online:2015-10-06 Published:2015-10-06
Contact: Lü Shi-Jie E-mail:jlmpcxlc@163.com

摘要/Abstract

摘要：

目的探讨胡桃醌对宫颈癌HeLa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HeLa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HeLa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HeLa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pAkt的表达。结果 MTT实验显示，与对照组比较，各时间点HeLa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，HeLa细胞Bcl-2,pAkt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HeLa细胞凋亡。

关键词: 胡桃醌, 磷脂酰肌醇3-激酶/Akt, HeLa细胞, 四甲基偶氮唑盐, Hoechst 33258染色, 免疫印迹法

Abstract:

[Abstract]：AIM:To Objective To explore the pro-apoptotic mechanism of juglone in human cervical cancer HeLa cells, and to study its associated signal transduction pathways. Methods Cultured HeLa cells were incubated with 30μmol/L juglone for 24,48 and 72 hours. The proliferation of HeLa cells was detected by methyl thiazolyl tetrazolium (MTT)assay. The cell apoptosis was detected by Hoechst 33258 staining and flow cytometry (FCM); The expressions of Bcl-2，Bax and Caspase-3,Akt,pAkt and PTEN were detected by Western blotting. Results MTT results showed that the HeLa cell growth was greatly inhibited in a time dependent manner (P<0.05,P<0.01) when compared with the control group. After treatment on HeLa cells for 4，8 and 12 hours, typical morphological changes of cell apoptosis such as karyopyknotic pyknic hyperfluorescence bolus，nuclear fragmentation and apoptotic body were observed by Hoechst 33258 staining. The early apoptosis ratio was increased significantly when compared with the control group. Western blotting results showed that the expression of Bcl-2 and pAkt decreased obviously，whereas those of Bax, Caspase-3 and Akt increased significantly in the juglone treatment group when compared with the control group. Conclusion Juglone may inhibit PI3K/Akt pathway, thereby promoting HeLa cells apoptosis.

Key words: Juglone, PI3K/Akt, HeLa cell, MTT, Hoechst 33258 staining, Western blotting

张巍张若文朱文赫李妍罗军赵行宇徐俊杰姜艳霞吕士杰*. 胡桃醌诱导HeLa细胞凋亡的机制[J]. 解剖学报, 2015, 46(5): 641-645.

ZHANG Wei ZHANG Ruo-wen ZHU Wen-he LI Yan LUO Jun ZHAO Xing-yu XU Jun-jie JIANG Yan-xia Lü Shi-Jie*. Effects of juglone on the proliferation and apoptosis of HeLa cells[J]. AAS, 2015, 46(5): 641-645.

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胡桃醌诱导HeLa细胞凋亡的机制

Effects of juglone on the proliferation and apoptosis of HeLa cells

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