目的 探讨槲皮素对缺氧人肝癌细胞HepG2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CoCl2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-1α mRNA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HepG2 增殖能力,这可能与槲皮素下调细胞中VEGF表达有关。
Abstract
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 and 72 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.
关键词
槲皮素 /
肝癌细胞 /
缺氧 /
缺氧诱导因子 /
血管表皮生长因子
Key words
quercetin /
hepatoma carcinoma cell /
hypoxia /
HIF-1α /
VEGF
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