AMPK-α/NF-κB信号转导通路在大肠癌多药耐药中的作用及白藜芦醇的干预机制

Effects of resveratrol on AMPK-α/NF-κB signaling pathway in multidrug resistance of colon cancer

目的:探讨白藜芦醇通过激活腺苷酸活化蛋白激酶α(AMPKα)抑制NF-κB/P-gp信号通路逆转大肠癌多药耐药的作用机制。方法:采用CCK-8法检测无毒浓度的白藜芦醇对人肠癌HCT116/OXA耐药细胞对奥沙利铂的逆转作用;细胞免疫荧光法检测白藜芦醇对p-AMPKα分布及表达的情况;Western blot法检测白藜芦醇对AMPKα-NF-κB/P-gp信号通路的调控作用;利用RNAi干扰技术沉默AMPKα基因,观察AMPKα介导的NF-κB/P-gp信号通路的调控作用。结果:白藜芦醇能够逆转大肠癌耐药细胞对奥沙利铂的耐药性,增加AMPKα磷酸化水平,抑制大肠癌细胞核内NF-κB的积累及其下游靶蛋白P-gp的表达;AMPKα活性降低能够增加NF-κB在细胞核内的积累及其下游靶蛋白P-gp的表达。结论:p-AMPKα表达下调可通过激活NF-κB信号通路并促进下游靶蛋白P-gp的表达以达到增加大肠癌细胞耐药的目的,而白藜芦醇通过激活AMPKα的表达发挥逆转大肠癌细胞多药耐药的作用。

Objective: To investigate the molecular mechanism of resveratrol on reversing multidrug resistance of colon cancer through inhibiting NF-κB/P-gp signaling pathway via AMPKα activation. Methods: The reversal effect of resveratrol at non-toxic concentrations on human colon cancer oxaliplatin-resistant cell HCT116/OXA was determined by CCK-8 assay. The distribution and expression of p-AMPKα were tested by immunocytochemical analysis. Regulation of resveratrol on AMPKα-NF-κB/P-gp signaling pathway was measured by Western blot. The si RNA was used to selectively knockdown AMPKα gene to observe NF-κB/P-gp signaling pathway mediated by AMPKα. Results: Resveratrol could reverse the oxaliplatin-resistant of colon cancer resistance cells, up-regulate AMPKα phosphorylation level and reduce the accumulation of NF-κB in the nuclear of colon cancer cells as well as inhibiting the expression of downstream target protein P-gp. Decreased AMPKα level could increase the accumulation of NF-κB in the nuclear and promote the expression of NF-κB downstream target protein P-gp. Conclusion: Down-regulated p-AMPKα level could enhance drug resistance of colon cancer cells by activating NF-κB signaling parhway and promoting the expression of downstream target protein P-gp, and resveratrol could reverse the multidrug resistance of colon cancer cells through AMPKα mediated NF-κB signaling parhway.