健脾解毒方介导JNK/SAPK信号通路调控人结肠癌细胞多药耐药

目的:探讨健脾解毒方(JPJD)介导JNK/SAPK信号通路对人结肠癌细胞多药耐药的调节机制。方法:人结肠癌细胞HCT8和多药耐药细胞HCT8/V常规培养,应用Western Blot检测健脾解毒方药物血清对P-糖蛋白(P-gp)以及c-Jun氨基端激酶(JNK)信号通路中c-Jun63/73磷酸化水平;RFQ-PCR检测MDR1 mRNA的表达;电感耦合等离子质谱法(ICP-MS)检测细胞内奥沙利铂(L-OHP)药物浓度的改变。结果:人结肠癌多药耐药细胞HCT8/V的P-gp和c-Jun的磷酸化表达明显高于HCT8细胞;与对照组比较,健脾解毒方药物血清作用48h后HCT8/V细胞的c-Jun的Ser63/73磷酸化、P-gp、MDR1 mRNA水平显著降低(P0.01),细胞组内L-OHP浓度明显增高(P0.01)。结论:健脾解毒方通过降低c-Jun的Ser63/73磷酸化,抑制JNK信号通路的激活,降低P-gp的表达,逆转HCT8/V细胞的多药耐药。

Effect of Jianpi Jiedu Recipe mediated JNK/SAPK signal transduction pathway on regulating multidrug resistance in human colon carcinoma cells

Objective: To investigate the mechanisms and reversal effects of Jianpi Jiedu Recipe on human colon carcinoma multidrug resistance(MDR) in HCT8/V cells.Methods: Human colon carcinoma cells were cultured conventionally.Expression of P-gp,c-Jun,and phosphorylated of c-Jun involved in JNK/SAPK signal transduction pathway were evaluated by Western blot in HCT8 and HCT8/V cells.After treatment with Jianpi Jiedu Recipe,expression of P-gp,phosphorylated c-Jun were evaluated by Western blot;MDR1 mRNA was analyzed by Real-time fluorescence quantitative PCR(RFQ-PCR);and the variation of intracellular concentration of L-OHP in HCT8/V cells was evaluated by the ICP-MS.Results: The expression of P-gp and phosphorylated c-Jun in HCT8/V cells were much higher than that in HCT8 cells.After treatment with Jianpi Jiedu Recipe for 48h,the activity of phosphorylated c-Jun at Ser63/73 was inhibited,and the expression of MDR1/P-gp was also up-regulation,compared with the control group(P<0.01).ICP-MS results showed that Jianpi Jiedu Recipe increased the concentration of L-OHP drugs in HCT8/V cells after treatment for 48h(P<0.01).Conclusion: Jianpi Jiedu Recipe reverse MDR in HCT8/V cell,which possibly through c-jun at Ser63/73 inhibiting the activation of JNK signal transduction pathway,ultimately regulate the expression of P-gp in MDR cells.