维生素K2通过降低D-双功能蛋白活性抑制肝癌的实验研究

目的探讨维生素K2 (VK2)抑制D-双功能蛋白(DBP)诱导肝癌细胞增殖的机制。方法将HepG2细胞分为过表达DBP组(DBP组)和空载体质粒对照组(Empty组)及敲低DBP组(siDBP组)和对照小干扰RNA组(siNC组),DBP组和siDBP组分别给予0~100μmol/L不同浓度的维生素K2处理。用DBP过表达质粒和干扰RNA分别转染HepG2细胞,Western-blot检测DBP表达的变化;CCK-8分析检测细胞增殖能力;放射免疫分析测定雌二醇(E2)的水平。结果 DBP组HepG2细胞的增殖显著高于Empty组(1.76±0.22 1.83±0.27,P <0.05),siDBP组细胞增殖显著低于siNC组(1.84±0.25 vs.1.77±0.21,P <0.05),DBP组E2水平低于Empty组(20.76±3.42 vs.17.23±2.65,P<0.05);DBP组给予VK2处理后,HepG2细胞的增殖呈剂量依赖性降低(P<0.05),E2的水平也呈剂量依赖性增加(P<0.05);但VK2不影响DBP的表达。结论 VK2抑制由DBP过表达引起的肝癌细胞增殖的机制,为VK2作为治疗肝癌的辅助分子提供了新的理论基础和实验依据。

Experimental study on the inhibition of liver cancer by vitamin K_2 by reducing the activity of D-bifunctional protein

Objective To explore the mechanism of vitamin K2(VK2) inhibiting the proliferation of hepatoma cells induced by D-bifunctional protein(DBP).Methods HepG2 cells were divided into overexpression DBP group(DBP group),empty vector plasmid control group(empty group),knockdown DBP group(siDBP group) and control small interfering RNA group(siNC group).The DBP group and sidbp group were treated with different concentrations of vitamin K2 from 0 to 100 μmol/L respectively.HepG2 cells were transfected with DBP over expression plasmid and interference RNA respectively.Western blot was used to detect the expression of DBP.CCK 8 analysis was used to detect cell proliferation.The level of estradiol(E2) was determined by radioimmunoassay.Results The proliferation of HepG2 cells in DBP group was significantly higher than that in Empty group(1.76±0.22 vs.1.83±0.27,P <0.05).The cell proliferation of siDBP group was significantly lower than that of siNC group(1.84±0.25 vs.1.77±0.21,P <0.05).The E2 level in the DBP group was lower than that in the Empty group(20.76±3.42 vs.17.23±2.65.P <0.05).After treatment with VK2 in the DBP group,the proliferation of HepG2 cells decreased in a dose-dependent manner(P <0.05).The level of E2 also increased in a dose-dependent manner(P <0.05);however,VK2 did not affect the expression of DBP.Conclusion The mechanism of VK2 inhibiting the proliferation of hepatoma cells induced by DBP overexpression provides a new theoretical basis and experimental basis for VK2 as an auxiliary molecule in the treatment of hepatoma.