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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
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。
[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.
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
Juglone / PI3K/Akt / HeLa cell / MTT / Hoechst 33258 staining / Western blotting
［1］Sugie S, Okamoto K, Rahman KM, et al. Inhibitory effects of plumbagin and juglone on azoxymethane-induced intestinal carcinogenesis in rats［J］. Cancer Lett, 1998,127(1):177-183.
［2］Chao SH, Greenleaf AL, Price DH. Juglone, an inhibitor of the peptidyl-prolyl isomerase Pin1, also directly blocks transcription［J］. Nucleic Acids Res, 2001,29(3):767-773.
［3］Varga Z, Bene L, Pieri C, et al. The effect of Juglone Oil the membrane potential and whole-cell K-currents of human lymphocytes［J］. Biochem Biophys Res Commun, 1996, 218(3):828-832.
［4］Xu HL, Yu XF, Qu SC, et al. Anti-proliferative effect of Juglone from Juglans mandshurica Maxim on human leukemia cell HL-60 by inducing apoptosis through the mitochondria-dependent pathway［J］. Eur J Pharmacol, 2010, 645(1-3):14-22.
［5］Jing X, Jun B, Xifeng Sh, et al. Proliferation inhibition of human cervical cancer HeLa cells by casticin in vitro［J］. Chinese-German Journal of Clinical Oncology，201，10(1) : 47-50.
［6］Malaguti C, Ciminiello P, Fattorusso E, et al. Caspase activation and death induced by yessotoxin in HeLa cells ［J］. Toxicol In Vitro, 2002,16(4):357-363.
［7］Zhang W, Liu A, Li Y, et al. Anticancer activity and mechanism of juglone on human cervical carcinoma HeLa cells［J］. Can J Physiol Pharmacol, 2012,90(11):1553-1558.
［8］Zhang W,Li Y,Luo J, et al. Proliferation inhibition and apoptosis induction of Juglone on human cervical cancer Caski cells［J］. Jouraal of Hygiene Research, 2014, 43(6): 959-962. (in Chinese)
张巍,李妍,罗军,等. 胡桃醌对宫颈鳞癌Caski 细胞增殖及凋亡作用的研究［J］. 卫生研究, 2014, 43(6): 959-962.
［9］Tan ML, Ooi JP, Ismail N, et al. Programmed cell death pathways and current antitumor targets［J］. Pharm Res, 2009, 26(7): 1547-1560.
［10］Xu Y, Li ZhX,Cao HL,et al.Effect of 3-methyladenine cisplatin induced-apoptosis of line hela cell［J］. Acta Anatomica Sinica, 2012, 43(1):59-62. (in Chinese)
徐冶, 李质馨, 曹慧玲, 等. 3-甲基腺嘌呤对顺铂诱导HeLa细胞凋亡的影响［J］.解剖学报, 2012, 43(1):59-62.
［11］Datta SR, Dudek H, Tao X, et al. Akt phosphorylation of BAD couples survival signals to the cell-intrinsic death machinery［J］. Cell, 1997,91(2): 231-241.
［12］Gu Ch,Xu ShL,Tang WW, et al. HeLa cell apoptosis inducedby quercetin and effects of caspase-3，caspase-8 activationon apoptosis［J］. Journal of Chinese Pharmaceutical Sciences, 2011, 46(8):595-599.
［13］Chen QP, Huang SS, Zhou WW, et a. Role of PI3K/Akt signal pathway in opening mito-KATP to inhibit the expression of FKN in rat myocardium microvascular endothelia cells exposed to hypoxia-reoxygenation［J］. Acta Anatomica Sinica, 2012, 43(6):778-783. (in Chinese)
陈秋萍,黄赛赛,周伟伟,等. PI3K/Akt在线粒体ATP敏感性钾通道开放抑制缺氧复氧大鼠心肌微血管内皮细胞FKN表达中的作用［J］. 解剖学报, 2012, 43(6):778-783.
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