Li Wei, Zhao Ying, Tao Bo, Zhang Ying
Department of Pharmacy, Harbin Commercial University, Harbin, 150076, China.
Chin J Integr Med. 2014 Oct;20(10):776-81. doi: 10.1007/s11655-014-1817-3. Epub 2014 Jun 13.
To investigate the effects of quercetin on Hedgehog (Hh) signaling in chronic myeloid leukemia KBM7 cells.
The KBM7 cells were treated with 50, 100 and 200 μmol/L quercetin for 48 h respectively. And then the trypan blue assay was used to examine the proliferative inhibition of quercetin. Apoptotic cells and cell cycle were measured by flow cytometry. The mRNA and protein expression were detected by quantitative real-time polymerase chain reaction (PCR) and Western blot, respectively.
Quercetin significantly inhibited KBM7 cell proliferation, induced cell apoptosis, and blocked cell cycle at G1 phase, which were in dose-dependent manners. The mRNA and protein expression of Smoothened and Glioma1 (Gli1), the members of Hh pathway decreased after treatment with quercetin. The Bcl-2 and Cyclin D1, targets of Hh signaling, also decreased after treatment with quercetin, respectively. Quercetin also could increase p53 and Caspase-3 expression. Bcr-abl mRNA copies decreased, but no changes of phosphorylated Bcr-abl and Bcr-abl proteins were observed, after treatment with quercetin.
Quercetin could inhibit Hh signaling and its downstream targets in the KBM7 cells. And it might be one of mechanisms of inducing apoptosis and inhibiting cell cycle by quercetin.
研究槲皮素对慢性髓性白血病KBM7细胞中刺猬信号通路(Hh)的影响。
将KBM7细胞分别用50、100和200μmol/L槲皮素处理48小时。然后用台盼蓝试验检测槲皮素的增殖抑制作用。通过流式细胞术检测凋亡细胞和细胞周期。分别用定量实时聚合酶链反应(PCR)和蛋白质免疫印迹法检测mRNA和蛋白质表达。
槲皮素显著抑制KBM7细胞增殖,诱导细胞凋亡,并将细胞周期阻滞在G1期,呈剂量依赖性。用槲皮素处理后,Hh信号通路成员Smoothened和胶质瘤相关癌基因1(Gli1)的mRNA和蛋白质表达降低。Hh信号通路的靶标Bcl-2和细胞周期蛋白D1在用槲皮素处理后也分别降低。槲皮素还可增加p53和半胱天冬酶-3的表达。用槲皮素处理后,Bcr-abl mRNA拷贝数减少,但未观察到磷酸化Bcr-abl和Bcr-abl蛋白的变化。
槲皮素可抑制KBM7细胞中的Hh信号及其下游靶标。这可能是槲皮素诱导细胞凋亡和抑制细胞周期的机制之一。