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目的观察槲皮素对肝癌HepG2细胞生长及对hTERT基因表达的影响。方法以台盼蓝拒染法计数肝癌细胞的生长抑制率,透射电镜从形态变化上了解凋亡的发生,流式细胞术检测细胞周期变化,western-blot、RT-PCR检测hTERT基因表达改变,PCR-TRAP法检测端粒酶活性。结果台盼蓝拒染法计数显示槲皮素抑制肝癌HepG2细胞增殖的作用明显,且呈浓度和时间依赖性,槲皮素处理48h后的Ic50为25.5μm。形态学检测显示出细胞凋亡的特征变化,流式细胞仪检测表明经10～20μm/L的槲皮素处理,肝癌HepG2细胞周期阻滞于G0/G1期,且HepG2细胞hTERT蛋白和mRNA表达降低,端粒酶活性受抑制。结论槲皮素能抑制肝癌细胞的生长,呈时间、剂量依赖性,能诱导HepG2细胞发生凋亡,其抑制增生与诱导凋亡的机制可能与下调hTERT基因表达,抑制端粒酶活性,破坏端粒稳定性有关。
Objective To observe the effects of quercetin on the growth of HepG2 cells and the expression of hTERT gene. Methods The inhibitory rate of hepatocellular carcinoma cells was counted by trypan blue exclusion method. The morphological changes were observed by transmission electron microscopy. The cell cycle was detected by flow cytometry. The expression of hTERT gene was detected by western-blot and RT-PCR. The telomerase activity was detected by PCR-TRAP. Results The results of trypan blue exclusion assay showed that quercetin significantly inhibited the proliferation of HepG2 cells in a concentration- and time-dependent manner. The Ic50 after 48 h of quercetin treatment was 25.5 μm. Morphological examination revealed the change of apoptosis. Flow cytometry showed that the cell cycle of HepG2 cells was arrested in G0 / G1 phase after treated with 10-20 μm / L quercetin, and the expression of hTERT protein and mRNA in HepG2 cells Decreased, telomerase activity was inhibited. Conclusions Quercetin can inhibit the growth of HepG2 cells in a time-and dose-dependent manner and induce apoptosis in HepG2 cells. Quercetin may inhibit the proliferation and induce apoptosis of HepG2 cells in a dose-dependent manner. The mechanism may be related to down-regulation of hTERT gene expression, inhibition of telomerase activity, Grain stability related.