黄连-大黄-肉桂复方及其组分对人肝癌SMMC-7721及HepG2细胞增殖的影响

目的观察黄连-大黄-肉桂复方及其组分配伍对人肝癌SMMC-7721及HepG2细胞增殖的影响,并从PI3K/AKT-FOXO3a通路探讨其作用的分子机制。方法体外培养人肝癌SMMC-7721及HepG2细胞,采用MTT法分别检测复方和复方中黄连、大黄、肉桂各自主要有效成分小檗碱、大黄素和桂皮醛不同配伍的肝癌细胞增殖;进一步以复方及有效组分最佳配伍(简称"组分")作用肝癌细胞24h,RT-qPCR检测PI3K、AKT和FOXO3a mRNA表达,Western blot检测PI3K p110、AKT、p-AKT、FOXO3a和p-FOXO3a蛋白表达。结果复方、小檗碱、大黄素和桂皮醛对肝癌细胞增殖具有抑制作用,并有较明显的量效和时效关系,小檗碱、大黄素和桂皮醛最佳配伍比例分别为44、23、46μmol/L;与对照组比较,复方和组分均能显著上调人肝癌SMMC-7721及HepG2细胞FOXO3a mRNA表达,PI3K mRNA表达也呈上调趋势,SMMC-7721细胞AKT mRNA表达显著下调,但组分使HepG2细胞AKT mRNA表达显著上调;复方和组分均可抑制PI3K p110、AKT及p-AKT蛋白表达,显著促进FOXO3a蛋白表达,对p-FOXO3a蛋白表达有抑制趋势。结论复方和组分均能抑制SMMC-7721、HepG2细胞增殖,其抗肝癌作用可能与PI3K/AKT的抑制和FOXO3a的激活有关。

Effects of Coptidis Rhizoma-Rhei Radix et Rhizoma-Cinnamomi Cortex Compound and Its Component on Proliferation of Hepatoma SMMC-7721 and HepG2 Cells

Objective To observe the effects of Coptidis Rhizoma-Rhei Radix et Rhizoma-Cinnamomi Cortex compound and its component compatibility on proliferation of hepatoma SMMC-7721 and HepG2 cells;To explore the action mechanism in the phosphatidylinositol 3-kinase(PI3K)-protein kinase B(AKT)-forkhead factor 3(FOXO3a)pathway.Methods The SMMC-7721 and HepG2 cells were cultured in vitro.MTT assay was used to determine the effect of Coptidis Rhizoma-Rhei Radix et Rhizoma-Cinnamomi Cortex compound and its main active component compatibility of berberine,emodin and cinnamaldehyde on the proliferation hepatoma cells,respectively.Furthermore,hepatoma cells were treated with the compound and the best compatibility for 24 hours.The mRNA expressions of PI3K,AKT and FOXO3a were detected by RTqPCR,and the protein expressions of PI3K p110,AKT,p-AKT,FOXO3a and p-FOXO3a were measured by Western Blot.Results The inhibitory effects of the compound,berberine,emodin and cinnamaldehyde on the proliferation of hepatoma cells were significant,with obvious dose-effect and time-effect relationship.The optimal compatibility ratio of berberine,emodin and cinnamaldehyde was 44,23,46μmol/L.Compared with the control group,both compound and component compatibility could significantly up-regulate the mRNA expression of FOXO3 a in SMMC-7721 and HepG2 cells,and also up-regulate the mRNA expression of PI3K;the mRNA expression of AKT in SMMC-7721 cells was significantly down-regulated.However,the component compatibility made the mRNA expression of AKT remarkably up-regulated in HepG2 cells;Compared with the control group,compound and component compatibility could inhibit the protein expressions of PI3K p110,AKT and p-AKT,notably promote the protein expression of FOXO3a,and reduce the protein expression of p-FOXO3a.Conclusion Both compound and component compatibility can inhibit the proliferation of SMMC-7721 cells and HepG2 cells.The anti-hepatocarcinoma effect may be related to the inhibition of PI3K/AKT and the activation of FOXO3a.