羟基喜树碱对人肺癌A549细胞自噬的影响

Effect of hydroxycamptothecin on autophagy of A549 lung cancer cells

目的探究羟基喜树碱(HCPT)对人肺癌A549细胞自噬的作用。方法体外进行肺癌A549细胞的培养,分别使用50、200和400μmol的HCPT处理,利用MTT法检测HCPT对细胞存活率的影响;共聚焦显微镜成像技术观察HCPT对细胞自噬活性的影响;Western blot检测HCPT对自噬相关蛋白LC3、p-mTOR表达的影响。结果 HCPT抑制A549细胞的活性;HCPT处理24 h后细胞质和细胞核周围CytoID绿色荧光强度增强;与对照组比较,HCPT治疗各组LC3II/I值升高,p-mTOR表达减少;且加入自噬抑制剂6-氨基-3-甲基嘌呤(3-MA)处理后的各组与相对应的HCPT治疗各组比较,A549细胞的增殖抑制增强。结论 HCPT能抑制A549细胞增殖并诱导其发生自噬;HCPT发挥抑制A549细胞增殖作用可能与自噬密切相关;抑制细胞自噬可能会增强HCPT的抗A549细胞增殖能力。

Objective To investigate whether autophagy is involved in the anticancer mechanisms of hydroxycamptothecin(HCPT)in human lung cancer A549cells in vitro.Methods Human non-small cell lung cancer cell line A549was cultured.In our study,cell viability was assessed by MTT assay,Cyto-ID staining was used for autophagy detection which was observed by laser scanning confocal microscope(LSCM),the expression of LC3II/I and p-mTOR were measured by Western blot.Results HCPT(50,200and400μmol)suppressed the viability of A549cells in a concentration-dependent manner.Moreover,HCPT increased autophagy level in the A549cells.The autophagy inhibitor3-MA caused a statistically significant increase in growth inhibition of the HCPTtreated A549cells.Conclusions HCPT induces autophagy in lung cancer A549cells in vitro.HCPT(50,200and400μmol)suppresses the viability of A549cells in a concentration-dependent manner and its induction of cell autophagy is related to the dosage.Autophagy inhibition combined with HCPT may be a better way for the treatment of lung cancer.