5-氟尿嘧啶诱导人肺腺癌A549细胞自噬  

5-fluorouracil induced autophagy in human lung adenocarcinoma A549 cells

作　　者：潘效红[1] 王娟[1] 张瑾锦[1] 闫苗苗[1] 于敏[1] 

机构地区：[1]滨州医学院药学院细胞工程教研室,山东烟台264003

出　　处：《解剖学报》2011年第3期374-377,共4页Acta Anatomica Sinica

基　　金：滨州医学院科技计划资助项目(BY2008KJ31);山东省高校科技计划资助项目(J10LC66)

摘　　要：目的探讨5-氟尿嘧啶(5-FU)是否诱导人肺腺癌A549细胞发生自噬。方法吖啶橙(AO)荧光染色和透射电镜观察自噬泡的变化;细胞免疫化学法测定LC3-Ⅱ的表达;流式细胞术及吖啶橙染色结合激光扫描共焦显微镜检测细胞凋亡;DNA电泳检测凋亡梯形条带的产生。结果吖啶橙荧光染色和透射电镜观察结果显示,5-氟尿嘧啶处理细胞后,细胞质中均出现明显的自噬泡;细胞免疫化学法的结果显示,5-氟尿嘧啶处理细胞后,细胞质中LC3-Ⅱ的表达明显升高,且3-甲基腺嘌呤(3-MA)能明显抑制LC3-Ⅱ的升高;流式细胞术、吖啶橙染色和DNA ladder结果表明,5-氟尿嘧啶处理细胞后,细胞未发生凋亡。结论低浓度(5μmol/L)的5-氟尿嘧啶能诱导A549细胞发生自噬而非凋亡,自噬特异性抑制剂3-MA能明显抑制细胞自噬的发生。Objective To study if 5-fluorouracil （5-FU） can induce human lung adenocarcinoma A549 cells＇ autophagy in order to provide theoretical basis for the tolerance of the chemotherapeutics. Methods Autophagic vacuole＇ s change was observed with acridine orange （AO） staining under fluorescent microscope and transmission election microscope （TEM） ; LC3-Ⅱ expression was tested with cellular immunochemistry; Apoptosis was detected by flow cytometry and AO staining with laser scanning confocal microscopy detection; Production of apoptosis＇ ladder-shaped stripe was tested with DNA cataphoresis. Results After being treated with 5-fluorouracil, the ceils cytoplasm appeared to have apparent autophagic vacuoles when observed with AO staining and TEM observations; The cells showed that LC3-Ⅱ expression was apparently increased and 3-MA significantly inhibited the increase of LC3-Ⅱ when observed with cellular immunoehemistry; The cell apoptosis did not occur when tested by flow cytometry, AO staining and DNA ladder. Conclusion Low concentration（51μmol/L） of 5-fluorouracil can induce A549 cells＇ autophagy rather than apoptosis and autophagic inhibitor 3-MA which has specificity can apparently inhibit the occurrence of autophagy.

关 键 词：5-氟尿嘧啶 肺腺癌 自噬 免疫细胞化学 流式细胞术 人 

分 类 号：R3[医药卫生—基础医学]