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神经节苷脂GM3对人宫颈癌HeLa细胞凋亡与血管内皮生长因子表达的影响

Effects of Monosialoganglioside-3 on Apoptosis and VEGF Expression in Human Cervical Cancer Cell Line HeLa

  • 摘要: 目的 研究外源性神经节苷脂GM3对人宫颈癌HeLa细胞增殖、凋亡和VEGF表达的影响。方法 不同浓度GM3干预HeLa细胞24 h后,MTT法检测细胞增殖变化,流式细胞学技术检测细胞凋亡率,RT-PCR技术检测细胞VEGF mRNA表达水平,Western blot技术检测细胞VEGF蛋白水平。结果(1)GM3浓度分别为2 、10、50和250 μmol/L时,HeLa细胞生长抑制率分别为9.8%、32.9%、50.7%和78.6%,半数抑制浓度(IC50)为49.8 μmol/L。(2)GM3浓度分别为10、50和250 μmol/L时,HeLa细胞凋亡率分别为(4.9±0.4)%、(15.1±0.3)%、(24.4±0.7)%。(3)当GM3浓度高于10 μmol/L时,HeLa细胞VEGF mRNA表达水平以及VEGF蛋白表达水平均显著下降(P<0.05)。结论 外源性神经节苷脂GM3能呈浓度依赖性抑制人宫颈癌HeLa细胞增殖,促进HeLa细胞凋亡,下调HeLa VEGF mRNA水平和蛋白水平,提示GM3可能通过调节肿瘤细胞凋亡和肿瘤血管生成而发挥双重抗肿瘤作用。

     

    Abstract: Objective To determine the effect of exogenous monosialoganglioside-3(GM3) on the proliferation, apoptosis and VEGF regulation in human cervical cancer cell line HeLa. Methods HeLa cells were treated with GM3 at different concentrations for 24h. MTT was used to detect cell proliferation and flow cytometry was applied to analyze cell apoptosis. RT-PCR was used to detect the expression level of VEGF mRNA and Western blot was used to detect the expression level of VEGF protein. Results (1) When GM3 concentrations were 2, 10, 50 and 250μmol/L, the inhibition rates of HeLa cells growth were 9.8%, 32.9%,50.7% and 78.6%. The IC50 was 49.8μmol/L. (2) When GM3 concentrations were 10, 50 and 250μmol/L,HeLa cells apoptosis rates were (4.9±0.4)%, (15.1±0.3)% and (4.4±0.7)%. (3) Comparing with the control group, when GM3 concentrations were higher than 10 mol/L, the expression level of VEGF mRNA and protein in HeLa cells were significantly decreased (P<0.05). Conclusion Exogenous GM3 could inhibit the proliferation and promote the apoptosis of HeLa cells, and down-regulate the VEGF mRNA and protein expression levels in HeLa cells in a dose-dependent manner. This may be considered as two mechanisms of anti-tumor effect of GM3 by modulating cell apoptosis and angiogenesis.

     

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