至真方调控Hedgehog/ABCG2信号途径改善大肠癌细胞的多药耐药性

ZZR reverses the multidrug resistance of colorectal cancer cells through Hedgehog/ABCG2 signaling pathway

[目的]探讨至真方醇提物(ZZR)对大肠癌多药耐药细胞HCT-8/5-FU细胞耐药性的改善作用,并从Hedgehog/ABCG2信号途径探讨至真方改善大肠癌多药耐药的可能作用机制。[方法]高效液相色谱法鉴定至真方醇提物中的5种主要成分;CCK-8法检测大肠癌多药耐药细胞HCT-8/5-FU的多药耐药性及至真方对其耐药性的改善作用;应用Real-Time PCR和western blot方法分别从基因和蛋白水平检测至真方对Hedghog/ABCG2信号途径的调节作用。[结果]HCT-8/5-FU细胞有明显多药耐药性,至真方醇提物干预HCT-8/5-FU细胞48h后,能增加其对不同化疗药物(5-FU、L-OHP、Taxol)的敏感性,且Hedgehog通路上关键因子-Gli1与ABCG2的mRNA相对表达量均明显下调(P<0.05),蛋白表达下降(P<0.05)。[结论]至真方醇提物能够有效改善HCT8/5-FU细胞多药耐药,其机制可能与抑制Hedgehog信号通路活性从而降低ABCG2蛋白表达有关。

[Objective]To investigate the effect of ZZR on the multidrug resistance of HCT-8/5-FU cells and to explore the mechanism of ZZR in multidrug resistance.[Methods]HPLC was used to quantify the five active ingredients in the alcohol extract of ZZR.The CCK-8 method was used to detect the multidrug resistance of HCT-8/5-FU.The real-time PCR and western blot were used to detect the regulation of Hedghog/ABCG2 signaling pathway by ZZR on mRNA and protein level.[Results]The HCT-8/5-FU cells had significant multidrug resistance,its IC50 for 5-FU,L-OHP and Taxol was significantly higher than that in the control group（P〈0.05）.The mRNA and protein expression of Gli1 and ABCG2 was significantly downregulated by ZZR in HCT-8/5-FU cells（P〈0.05）.[Conclusion]ZZR can effectively improve the multidrug resistance of HCT8/5-FU cells.The mechanism may be related to the inhibition of Hedgehog signaling pathway and the expression ABCG2.