槲皮素对缺氧人肝癌细胞HepG2增殖及HIF-1α、VEGF的影响

Effects of quercetin on proliferation of hypoxia induced hepatoma carcinoma cell HepG2 and expressions of HIF-1α 、VEGF

目的探讨槲皮素对缺氧人肝癌细胞Hep G2增殖能力、HIF-1α以及VEGF表达的影响及其可能机制。方法实验的细胞分为正常对照组、缺氧组、槲皮素+缺氧组,采用Co Cl2法诱导缺氧环境。采用MTT法检测各组细胞增殖能力,实时定量PCR及Western blot检测各组细胞缺氧诱导因子HIF-1α和VEGF mRNA及蛋白表达。结果与正常对照组比较,缺氧组细胞在24、48、72 h增殖能力降低(P<0.05),与缺氧组比较,槲皮素+缺氧组细胞在24、48、72 h增殖能力显著降低(P<0.05),正常组在24、48、72 h的光密度值分别为0.640±0.016、1.287±0.025、1.738±0.027,缺氧组的光密度值分别为0.551±0.021、0.851±0.028、1.268±0.018,槲皮素+缺氧组的光密度值分别为0.435±0.023、0.717±0.013、0.984±0.037。与正常对照组比较,缺氧组细胞HIF-1α、VEGF的mRNA及蛋白表达上调(P<0.05),而槲皮素可下调缺氧细胞中VEGF mRNA及蛋白表达,对HIF-1αmRNA及蛋白表达无影响(P>0.05):正常对照组HIF-1α、VEGF的mRNA表达均为1.000±0.000,蛋白表达分别为0.616±0.016、0.831±0.034,缺氧组HIF-1α、VEGF的mRNA表达分别为2.298±0.031、2.065±0.090,蛋白表达分别为1.385±0.047、2.022±0.076,槲皮素+缺氧组HIF-1α、VEGF的mRNA表达2.326±0.027、0.990±0.077,蛋白表达分别为1.406±0.029、1.137±0.054。结论槲皮素可抑制缺氧肝癌细胞Hep G2增殖能力,这可能与槲皮素下调细胞中VEGF表达有关。

Objective To explore the effect of quercetin on the proliferation and expression of HIF-1α and VEGF,and the mechanisms in hypoxia hepatoma carcinoma cell HepG2.Methods The cells were divided into normal control group,hypoxia group and hypoxia combined with quercetin group.A hypoxia model was established using cobalt chloride（CoCl2）.The proliferation of cells was determined by MTT assay.The mRNA expression and protein expression of HIF-1α and VEGF were determined by real-time quantification PCR and Western blot.Results Compared with the normal control group,the proliferation of the hypoxia group was decreased at 24,48 and 72 h（P〈0.05）,while the proliferation in hypoxia combined with quercetin group was decreased at 24,48 and72 h（P〈0.05） compared with the hypoxia group.The optical density was 0.640 ± 0.016、1.287 ± 0.025 and 1.738 ± 0.027 respectively in normal control group,0.551 ± 0.021、0.851 ± 0.028 and 1.268 ± 0.018 in hypoxia group,and 0.435 ± 0.023、0.717 ±0.013 and 0.984 ± 0.037 respectively in hypoxia combined with quercetin group.The mRNA and protein expressions of HIF-1α and VEGF in hypoxia group were increased（P〈0.05） compared with normal control group.Quercetin down-regulated the mRNA and protein expressions of VEGF（P〈0.05） in hypoxia cells,but had no effect on the mRNA and protein expression of HIF-1α（P〈0.05）.The mRNA expressions of HIF-1α and VEGF were both 1.000 ± 0.000 and the protein expressions were 0.616 ± 0.016 and 0.831 ±0.034 in normal control group.The protein expressions were 1.385 ± 0.047 and 2.022 ± 0.076 in the hypoxia group,but were 1.406± 0.029 and1.137 ± 0.054 in the hypoxia combined with quercetin group.Results Quercetin can inhibit the proliferation of hepatoma carcinoma cell HepG2 under hypoxia conditions,which may be related to down-regulated expressions of VEGF.