染料木黄酮对喉癌Hep-2细胞系增殖和凋亡的影响

目的　探讨染料木黄酮（genistein，Gen）对人鳞状细胞喉癌Hep-2细胞系增殖和凋亡的影响。方法　用CCK-8法检测不同浓度Gen对Hep-2细胞系增殖的影响；用实时荧光定量聚合酶链反应（Real-Time PCR）检测Gen对Hep-2细胞系血管内皮生长因子受体-3（vascular endothelial growth factor receptor-3，VEGFR-3）mRNA表达的影响；用流式细胞术检测Gen单独及联合VEGFR-3特异性阻滞剂MAZ51对Hep-2细胞系凋亡的影响；Western blot检测各组AKT蛋白表达及其磷酸化水平。结果　Gen可抑制Hep-2细胞系增殖，且抑制作用呈剂量依赖性；Gen能抑制Hep-2细胞系的VEGFR-3 mRNA表达，抑制率为32%；单用Gen和MAZ51均能诱导Hep-2细胞系凋亡，二者联合应用对Hep-2细胞系的凋亡诱导作用明显增强；Gen与MAZ51均能抑制AKT第308位苏氨酸（Thr308）及第473位丝氨酸（Ser473）磷酸化激活，使p-AKT-Thr308和p-AKT-Ser473减少，两种药物合用抑制率更高。结论　Gen可通过抑制VEGFR-3表达阻止AKT磷酸化激活，并发挥抑制喉癌细胞增殖和诱导其凋亡的抗癌作用。

Impact of genistein on proliferation and apoptosis of human laryngeal cancer Hep-2 cell

OBJECTIVE To investigate the influence of Genistein （Gen） on proliferation and apoptosis of Hep-2 cell. METHODS The proliferation of Hep-2 cell after treated with various concentrations Gen was detected. The changes of VEGFR-3 mRNA were detected by real-time polymerase chain reaction （Real-Time PCR） in Hep-2 cells treated by Gen. Using flow cytometry （FCM）, the variation of cell apoptosis rate after treating Gen alone or Gen combined with MAZ51（a VEGFR-3-specific blocker） were measured. AKT proteins expression and phosphorylation were determined by Western blot. RESULTS CCK-8 assay showed the proliferation inhibition on Hep-2 cells had a dose-dependent effect treated by Gen. Gen could inhibit the expression of VEGFR-3 mRNA, and the inhibition rate was 32%. FCM showed that Gen and MAZ51 can both induce apoptosis in Hep-2 cells, combination Gen with MAZ51 could induce apoptosis more. The phosphorylations and expressions of the 308th threonine and the 473th serine of AKT were inhibited by Gen alone or MAZ51 alone, and the inhibiting effect was enhanced because using both of them. CONCLUSION Gen can inhibit VEGFR 3 expression, stop AKT phosphorylation, inhibit proliferation and induce apoptosis of laryngeal cancer cells, which plays an anti-tumor role.