摘要
研究非甾类抗炎药布洛芬对肝癌细胞增殖和β-catenin信号通路的影响,探讨布洛芬影响肝癌细胞增殖的可能机制。体外培养HepG2细胞,MTT法检测不同浓度布洛芬分别作用24、48和96h后细胞增殖情况;应用RT-PCR检测布洛芬处理HepG248h后β-catenin、cyclinD1和c-myc转录水平的变化;western blotting检测布洛芬对HepG2中β-catenin1蛋白表达量的影响;免疫细胞化学法观察布洛芬处理组HepG2细胞β-catenin的亚细胞定位。结果表明0.7mmol/L的布洛芬处理细胞48h后能明显见到细胞增殖受到抑制,抑制率达到53.8%,且抑制作用有时间、剂量依赖性;经布洛芬处理后细胞的β-catenin、cyclinD1和c-myc转录也受到显著抑制;布洛芬处理组细胞β-catenin蛋白表达低于对照组;布洛芬处理后β-catenin在胞核中定位减少,胞浆定位增多。因此,布洛芬能抑制肝癌细胞的增殖,且随着处理时间和药物浓度的增加,其抑制作用逐渐增强,其抑制作用可能与调控β-catenin信号通路,影响β-catenin的表达和定位,调节下游相关靶基因的转录有关。
It aimed to investigate the effects of ibuprofen,a nonsteroids anti-inflammatory drug（NAID）,on proliferation and β-catenin signaling pathway of human liver cancer cell line HepG2, and to discuss its possible mechanism of anti-liver-cancer as well. HepG2 cells were cultured in vitro and the cell proliferation was assessed by MTT assay by treatment of ibuprofen with various concentrations for 24,48 and 72 hours respectively. RT-PCR tests was used to analyze the transcription of β-catenin,cyclinD1 and c-myc after treatment of ibuprofen for 48 hours. Western blotting assay was introduced to investigate the influence of expression of β-catenin protein immunocytochemistry. Results indicated proliferation of HepG2 cell was inhibited by treatment of ibuprofen in a time-dependent and dose-dependent manner. ILT-PCT tests showed that the transcription of β-catenin,cyclin Dl,c-myc was inhibited by ibuprofen. Western blotting assay showed that the expression of β-catenin protein in ibuprofen-treated cells was lower than that of control cells. Therefore,ibuprofen could inhibit the proliferation of liver neoplasms cells HepG2 in a timedependent and time dose-dependent manner,which may be associated with regulation of the β-catenin signaling pathway, alteration of β-catenin expression,and location and also inhibition of downstream target gene transcription.
出处
《生物技术通报》
CAS
CSCD
北大核心
2009年第2期132-137,共6页
Biotechnology Bulletin
基金
国家自然科学基金(30771925)
