至真方调控Hedgehog信号通路对HCT-8/VCR多药耐药的影响

The effect and mechanism of Zhizhen Recipe on multidrug resistance in human colorectal cancer cell lines through hedgehog signaling pathway

[目的]探讨至真方含药血清对人大肠癌耐长春新碱细胞株HCT-8/VCR多药耐药性的逆转作用,及对Hedgehog信号通路中Smo、Gli1表达的影响。[方法]CCK-8法测定细胞多药耐药性及存活率;Real-time PCR及Western blot分别从基因及蛋白水平检测P-gp和Hedgehog信号通路中Smo、Gli1的表达。[结果]至真方含药血清作用24h、48h后,HCT-8/VCR生存率明显降低;至真方含药血清作用24h后,HCT-8/VCR细胞P-gp(P-Glycoprotein)、Smo、Gli1基因及蛋白表达均降低,与阴性对照组相比差异有统计学意义(P<0.05),与阳性对照组相比差异无统计学意义(P>0.05)。[结论]至真方对HCT-8/VCR的逆转作用可能与其抑制Hedgehog信号通路,下调该通路相关蛋白表达进而下调P-gp表达,增加HCT-8/VCR对化疗药物的敏感性有关。

[Objective]To investigate the effect of serum containing Zhizhen Recipe on multidrug resistance（MDR）in human colorectal cancer cell lines HCT-8/VCR cells,and the influence on Smo and Gli1 expression in hedgehog signaling pathway.[Methods]Drug resistance of HCT-8/VCR cells and apotosis rate were detected with CCK-8.Gli1,Smo and P-gp protein levels were detected with Western blot and Real-time PCR.[Results]After treated with serum containing Zhizhen Recipe for 24 hand 48h,the survival rate of HCT-8/VCR was significantly decreased.The results of Western blot and Real-time PCR showed decreased expression of P-gp,Gli1 and Smo after treated for 24h（P0.05）.However,it was not statistically different compared to positive control group（P0.05）.[Conclusion]Serum containing Zhizhen Recipe can reverse the MDR of colon cancer cells.The possible mechanism might be that serum containing Zhizhen Recipe inhibited the hedgehog signal pathway and down regulated the expression of Gli1,Smo on gene and protein level,and finally improved the sensitivity of HCT-8/VCR cells to chemotherapy drugs.