黄芩苷对缺氧三阴性乳腺癌干细胞增殖及HIF-1α、IGF-Ⅱ、L-selectin的影响

Effects of baicalin on proliferation of hypoxic triple-negative breast cancer stem cells and on HIF-1α,IGF-Ⅱ,and L-selectin

目的研究黄芩苷对缺氧乳腺癌干细胞MDA-MB-231增殖及低氧诱导因子-1α(hypoxia inducible factor-1,HIF-1α)、胰岛素样生长因子-Ⅱ(insulin-like growth factor,IGF-Ⅱ)、L-选择素(L-selectin)的影响。方法四甲基偶氮唑蓝法检测不同浓度(12.5、25.0、50.0、100.0、200.0 mg/L)黄芩苷对缺氧乳腺癌干细胞MDA-MB-231的细胞生长抑制率的影响,筛选出合适的黄芩苷浓度。将缺氧乳腺癌干细胞MDA-MB-231随机分为10组:对照组、BL组(黄芩苷低浓度50.0 mg/L)、BM组(黄芩苷中浓度100.0 mg/L)、BH组(黄芩苷高浓度200.0 mg/L)、DDP组(顺铂)、M组(2-甲氧雌二醇)、BL+M组、BM+M组、BH+M组、DDP+M组。RT-PCR法检测HIF-1α、IGF-Ⅱ、L-selectin mRNA的表达,Western blot检测HIF-1α、IGF-Ⅱ、L-selectin蛋白的表达。结果与对照组比较,DDP组细胞生长抑制率明显升高(P<0.05),不同浓度黄芩苷的细胞生长抑制率均明显升高(P<0.05)。与黄芩苷200.0 mg/L比较,其他浓度黄芩苷的细胞生长抑制率均明显降低(P<0.05),DDP组细胞生长抑制率明显升高(P<0.05)。与对照组比较,其余9组组HIF-1α、IGF-Ⅱ、L-selectin蛋白和mRNA表达均明显降低(P<0.05)。与BL组比较,BM组、BH组、BL+M组HIF-1α、IGF-Ⅱ、L-selectin蛋白和mRNA表达均明显降低(P<0.05)。与BM组比较,BM+M组HIF-1α、IGF-Ⅱ、L-selectin蛋白和mRNA表达均明显降低(P<0.05)。与BH组比较,BH+M组HIF-1α、IGF-Ⅱ、L-selectin蛋白和mRNA表达均明显降低(P<0.05)。结论黄芩苷能抑制缺氧乳腺癌干细胞的增殖,可能通过调节HIF-1α/IGF-Ⅱ信号通路及L-selectin的表达抑制乳腺癌干细胞的增殖。

Objective To study the effects of baicalin on proliferation of hypoxic breast cancer cells MDA-MB-231,and on hypoxia-inducible factor-1α(HIF-1α),insulin-like growth factor-Ⅱ(IGF-Ⅱ)and L-selectin expression.Methods MTT assay was employed to detect inhibiting effect of different concentrations of baicalin(12.5,25.0,50.0,100.0,200.0 mg/L)on growth of hypoxic breast cancer stem cells MDA-MB-231,therefore identifying the appropriate baicalin concentration.The hypoxic breast cancer stem cells MDA-MB-231 were randomly divided into 10 groups:control group,BL group(baicalin low concentration 50.0 mg/L),BM group(baicalin medium concentration 100.0 mg/L),BH group(baicalin high concentration 200.0 mg/L),DDP group(cisplatin),M group(2-methoxyestradiol),BL+M group,BM+M group,BH+M group and DDP+M group.RT-PCR was employed to detect the expression of mRNA in HIF-1α,IGF-Ⅱand L-selectin.Western blot was applied to detect the protein expression of HIF-1α,IGF-Ⅱand L-selectin.Results Compared with the control group,the cell growth inhibition rate of DDP group and the cell growth inhibition rates at different concentrations of baicalin significantly increased(P<0.05).Compared with 200.0 mg/L baicalin,the cell growth inhibition rates of other concentrations of baicalin significantly decreased(P<0.05),while the cell growth inhibition rate of DDP group significantly increased(P<0.05).Compared with the control group,the protein and mRNA expression of HIF-1α,IGF-Ⅱand L-selectin in the other 9 groups significantly decreased(P<0.05).Compared with the BL group,the protein and mRNA expression levels of HIF-1α,IGF-Ⅱand L-selectin in the BM,BH and BL+M groups significantly decreased(P<0.05).Compared with the BM group,the protein and mRNA expression levels of HIF-1α,IGF-Ⅱand L-selectin in the BM+M groups significantly decreased(P<0.05).Compared with the BH group,the protein and mRNA expression levels of HIF-1α,IGF-Ⅱand L-selectin in the BH+M group significantly decreased(P<0.05).Conclusion Baicalin can inhibit the proliferation of hypoxic breast cancer stem cells.It may inhibit proliferation of breast cancer stem cells by regulating HIF-1α/IGF-Ⅱsignaling and L-selectin expression.