中山大学学报（医学科学版）2013，Vol.34Issue(1)：75-82,8.
漆黄素通过调节Apaf-1,ERK和COX-2信号通路诱导人宫颈癌细胞凋亡
Fisetin Simultaneously Targets Apaf-1, ERK, and COX-2 Signaling Leading to Growth Inhibition and Apoptosis in Human Cervical Carcinoma Cell In Vitro
摘要
Abstract
[Objective] To identify the molecular mechanisms by which fisetin inhibited cell proliferation and induced apoptosis in Hela cells. [Methods] Hela cells were treated with fisetin, and the effect of fisetin on cell growth and apoptosis was observed by MTT, PI, and Annexin V. The expression level and activity changes of apoptotic key proteins, caspase, and Apaf-1, the phosphorylated level of ERK1/2, and the expression of COX-2 and PGE2 were detected by Western blot, RT-PCR, immunofluorescence imaging, respectively. The inhibition by siRNA or inhibitors was used to confirm the function of above key proteins in fisetin-mediated antitumor effects. [Results] Fisetin, with a concentration of 80-200 μmol/L, inhibited the proliferation at a rate of 20% to 48% and with a concentration of 100-200 μmol/L, induced apoptosis of Hela cell. It increased the activities of Caspase-3 and Caspase-9 at ranges of 20% ～ 50% and 22%～ 38%, respectively, induced Apafl protein expression and cytochrome-C release. With the increase of the dosage, Fisetin inhibited the phosphorylation of ERK1/2 protein of the ERK MAPK signaling pathways. Combined with the ERK inhibitor or siRNA, it suppressed the proliferation at a rate of 60% to 70% and COX-2 protein expression of the Hela cell. Also, it reduced PGE2 production to 1800 pg/mL-2600 pg/mL and abrogated the binding of p300, NF-kB, p50, and p65 to COX-2 promoter. Compared with the control groups, these differences were significant (P < 0.05). [Conclusions] Fisetin induced apoptosis and proliferation inhibition by modulating the Apaf-1, ERK, and COX-2-dependent mechanisms in Hela cell.关键词
漆黄素/caspase/Apaf-1/ERK/COX-2/宫颈癌Hela细胞Key words
fisetin/ caspase/ Apaf-1/ ERK/ COX-2/ cervical cancer cell分类
医药卫生引用本文复制引用
刘立群,郭微,余文丹,游泽山..漆黄素通过调节Apaf-1,ERK和COX-2信号通路诱导人宫颈癌细胞凋亡[J].中山大学学报（医学科学版）,2013,34(1):75-82,8.基金项目
广东自然自然科学基金(S2011010003522) （S2011010003522）
