【摘要】 目的研究磷酸腺苷激活的蛋白激酶(AMPK)活化程度对人鼻咽癌(NPC)细胞放射敏感性的影响。方法应用AMPK活化抑制剂Compound C(CC)处理人NPC CNE-2细胞1h后,给予X线照射,48h后Western blot检测总AMPK(tAMPK)、磷酸化AMPK(p-AMPK)及自噬标志物MAPlLC3的变化;透射电子显微镜检测自噬小体形成情况。经不同药物干预后的CNE-2细胞给予X线照射48h后四甲基偶氮唑盐(MTT)和流式细胞技术检测两组细胞活力和凋亡变化。结果 CNE-2细胞经CC干预后t-AMPK未发生明显变化(P>0.05),而活化状态的AMPK,即p-AMPK表达显著降低(P<0.01),且自噬标志物MAPlLC3的表达量显著降低(P<0.05),自噬小体数量明显减少(P<0.05);CNE-2细胞经CC干预后进行X线照射可明显抑制其增殖并促进细胞凋亡(P<0.05)。结论 CC可通过抑制细胞自噬来增强放射线对NPC CNE-2细胞增殖抑制和凋亡促进作用,从而增强NPC的放射敏感性。更多还原

【Abstract】 Objective To investigate the effect of Amp activated protein kinase(AMPK)activity in radiosensitivity of human nasopharyngeal carcinoma cells.Methods Human nasopharyngeal carcinoma cells,CNE-2,were treated with AMPK inhibitor,Compound C(CC),for 1h.Then cells were explored in X-ray.The expression of total AMPK(t-AMPK),phosphorylation AMPK(pAMPK),and MAPlLC3 were detected by Western blot.The number of autophagosomes were observed and calculated by transmission electron microscope(TEM).Cells processed with CC were explored in X-ray.MTT assay was used to detect the difference of two groups in cell proliferation.Cell apoptosis were assayed by flow cytometry.Results The expression of p-AMPK in CC group cells were significantly downregulated compared to the negative control group cells(P<0.01),while no significant change of tAMPK expression were found(P>0.05).The expression of MAPlLC3 and the number of autophagosomes in CC group cells were significantly decreased compared to the control group cells(P<0.05).Correspondingly,the cell proliferation rate in CC group was lower than in control group,and the percent of apoptosis cells was higher in CC group than in control group(P<0.05).Conclusion Suppression of AMPK activity could inhibited autophagy induced by decreasing the degree of p-AMPK,then enhanced the effect of proliferation inhibition and apoptosis promotion in CNE-2cells.The AMPK inhibitor,CC,can serve as an effective assistant treatment of radiotherapy for nasopharyngeal carcinoma.更多还原