调节自噬对贝伐单抗诱导的肺癌A549细胞凋亡的影响  

Inhibition of autophagy enhances anticancer effects of bevacizumab in non-small cell lung caner

作　　者：董新伟[1] 温丰标[2] 银瑞[1] 卢万里[1] 吴恺[2] 杨洋[2] 赵松[2] 

机构地区：[1]南阳市中心医院胸外科,473009 [2]郑州大学第一附属医院胸外科河南省高等学校临床医学重点学科开放实验室郑州市胸部肿瘤重点实验室

出　　处：《中华实验外科杂志》2014年第2期336-337,共2页Chinese Journal of Experimental Surgery

基　　金：河南省高校科教创新团队资助项目（13IRTSTHN011）

摘　　要：目的 观察白噬在血管抑制药物贝伐单抗诱导的肺癌A549细胞凋亡中的作用.方法 设置空白对照组、贝伐单抗（16μmol/L）组、3-甲基腺嘌呤（3-MA,8μmol/L）组和3-MA＋贝伐单抗组4个实验组.收集各实验组细胞组,膜联蛋白Ⅴ-异硫氰酸荧光素（Annexin Ⅴ-FITC）和单丹磺酰戊二胺（MDC）染色后,荧光显微镜下定性观察荧光表达;流式细胞术定量检测细胞凋亡、Westernblot检测自噬相关蛋白Beclin 1和微管相关蛋白1轻链3（LC3）的表达.结果 （1）作用48 h,贝伐单抗对A549细胞的半数抑制浓度（IC50）为16 μmol/L,3-MA对A549细胞抑制率10％的浓度（IR10）为8μmol/L; （2）与单用贝伐单抗[（43.92±1.38）％]比较,3-MA联合贝伐单抗显著增加了细胞凋亡率[（87.46±5.97）％,P＜0.01],Beclin l和LC3的表达显著下调（P＜0.05）.结论 3-MA可能是通过抑制白噬信号通路,显著增加了贝伐单抗诱导的肺癌细胞凋亡.Objective To study the role of anti-proliferation effect of bevacizumab on autophagy inhibitor on non-small cell lung caner （NSCLC）.Methods The lung cencer cells A549 cells were treated with bevacizumab （Beva,16 μmol/L） and/or 3-methyladenine （3-MA,8 μmol/L）.Flow cytometry was performed to examine the cell apoptosis rate treated with annexin Ⅴ-fluoresceine isothiocyanate （FITC）/propidium iodide （PI） double staining; Cells stained by monodansylcadaverine （MDC） were observed qualitatively under a fluorescence microscope.Western blotting assay was used to investigate autophagy-related Beclin 1 and microtubule-associated protein 1 light chain 3 （LC3） changes that occurred in the course of treatment.Results The flow cytometry indicated that the respective apoptosis rate of Control,Beva （16 μ mol/L）,3-MA （8 μmol/L） and Beva ＋ 3-MA group were （3.45 ± 0.28） %,（43.92 ± 1.38） %,（7.63 ± 0.78） % and （87.46 ± 5.97） %.The autophagic vacuoles in Beva group were obviously increased,but the phenomenon was inhibited after combination with 3-MA.The expression of Beclin 1 and LC3 was clearly up-regulated in Beva group,while the degree of expression in combinition group dec reased significantly when compared with treatment with Beva alone.Conclusion Beva can induce the apoptosis and increase the autophagy in A549 cells,which could be reduced after co-treated with 3-MA.

关 键 词：血管生成抑制剂 自噬 肺癌 凋亡 

分 类 号：R734.2[医药卫生—肿瘤]