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目的:探讨Hedgehog(Hh)信号通路与人结肠腺癌细胞多药耐药的关系。方法:以结肠腺癌敏感细胞株HCT-8和多药耐药细胞株HCT-8/VCR为研究对象,采用Hh信号通路激动剂SAG及抑制剂GANT61进行干预。实验分为4组:HCT-8组、HCT-8/VCR组、HCT-8+SAG组(3nmol/L SAG干预48h)以及HCT-8/VCR+GANT61组(5μmol/L GANT61干预48h)。采用CCK-8(cell counting kit-8)法检测细胞耐药性及存活率。实时荧光定量PCR和蛋白质印迹法分别从基因和蛋白水平检测P-糖蛋白(P-glycoprotein,P-gp)及Hedgehog信号通路相关蛋白Smo和Gli1的表达。结果:HCT-8/VCR组Smo、Gli1和P-gp基因及蛋白表达高于HCT-8组(P<0.05);HCT-8+SAG组24和48h时的细胞存活率及Smo、Gli1、P-gp基因和蛋白表达水平均高于HCT-8组(P<0.05);HCT-8/VCR+GANT61组24和48h时的细胞存活率及Smo、Gli1、P-gp基因和蛋白表达均低于HCT-8/VCR(P<0.05)。结论:人结肠腺癌细胞多药耐药性与Hh信号通路异常激活有关。
Objective: To investigate the relationship between Hedgehog signaling pathway and multidrug resistance in human colon adenocarcinoma cells. Methods: Human colon adenocarcinoma cell line HCT-8 and multidrug-resistant cell line HCT-8 / VCR were used as experimental subjects, and HH signaling pathway agonist SAG and inhibitor GANT61 were used for intervention. The experiment was divided into 4 groups: HCT-8 group, HCT-8 / VCR group, HCT-8 + SAG group (48h for 3nmol / L SAG intervention) and HCT-8 / VCR + GANT61 group (48h for intervention with 5μmol / L GANT61). Cell chemoresistance and survival rate were detected by CCK-8 (cell counting kit-8). The expression of P-glycoprotein (P-gp) and Hedgehog signaling pathway-related proteins Smo and Gli1 were detected by real-time fluorescence quantitative PCR and Western blot respectively. Results: The mRNA and protein expressions of Smo, Gli1 and P-gp in HCT-8 / VCR group were significantly higher than those in HCT-8 group (P <0.05) The expression of P-gp gene and protein in HCT-8 / VCR + GANT61 group was significantly higher than that in HCT-8 group (P <0.05). The cell viability and the expression of Smo, Gli1 and P-gp Lower than HCT-8 / VCR (P <0.05). Conclusion: Multidrug resistance of human colon adenocarcinoma cells is related to abnormal activation of Hh signaling pathway.