至真方对人大肠癌多药耐药细胞株HCT-8/VCR中核因子-κB及P糖蛋白表达的影响

Effect of Zhizhen recipe on expression of NF-κB and P-gp in human colorectal cancer line HCT-8/VCR

[目的]从核因子-κB(NF-κB)途径研究至真方对大肠癌多药耐药(MDR)细胞株———人大肠癌细胞株及长春新碱细胞株(HCT-8/VCR)的逆转作用及P糖蛋白(P-gp)表达的影响。[方法]CCK-8法测定HCT-8/VCR的MDR性及至真方含药血清对HCT-8/VCR细胞株的逆转作用;Real-time PCR及Western blot检测NF-κB/p65、MDR1mRNA,NF-κB/p65、P-gp蛋白的表达。[结果]至真方含药血清可明显抑制HCT-8/VCR细胞生长。不同浓度至真方含药血清作用24h后,HCT-8/VCR细胞NF-κB/p65、MDR1mRNA及NF-κB/p65、P-gp蛋白表达均降低,且呈浓度依赖性,与阴性对照组相比差异均有统计学意义(P<0.05)。[结论]至真方对人大肠癌MDR细胞株HCT-8/VCR的逆转作用可能与其下调NF-κB、P-gp的表达相关。

[Objective]To investigate the effect of Zhizhen recipe in reversing multidrug resistance and the impact to the expression of P-gp of human colorectal cancer cell line HCT-8/VCR in NF-κB pathway.[Methods]The ability of multidrug resistance and the reversal effect of Zhizhen recipe contained serum were detected by CCK-8 assay.The expression of NF-κB/p65,MDR1 mRNA and NF-κB/p65,P-gp protein in HCT-8/VCR cell line were measured by Real-time PCR and Western blot.[Results]HCT-8/VCR was a kind of multidrug resistant cell line,and the drug-serum distinctly inhibited proliferation of HCT-8/VCR.After persistent treatment for 24 hours with different volume fractions of Zhizhen recipe contained serum,the expression of NF-κB/p65,MDR1 mRNA significantly decreased as well as NF-κB/p65 and P-gp protein.The level of their expression was volume-dependent,and obviously lower than negative control group（P〈0.05）.[Conclusion]Zhizhen recipe could reverse the multidrug resistance of HCT-8/VCR.The mechanism may be related to its effect of down-regulation of NF-κB and P-gp.