Inhibition of autophagy augments 5-fluorouracil chemotherapy in human colon cancer in vitro and in vivo model
Inhibition of autophagy augments 5-fluorouracil chemotherapy in human colon cancer in vitro and in vivo model
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DOI：
10.1016/j.ejca.2010.02.021
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发表时间：
2010-07-01
影响因子：
8.4
通讯作者：
Kuwano, Hiroyuki
中科院分区：
文献类型：
--
作者：
Li, Jie;Hou, Ni;Kuwano, Hiroyuki
尽管基于5-氟尿嘧啶（5-FU）的辅助化疗广泛用于结直肠癌的治疗，但仍需要探索新的治疗策略。据报道，自噬与癌症广泛相关。然而，自噬的功能尚不完全清楚。在本研究中，使用 MTT 测定、DNA 片段化测定、Hoechst 33342 染色和 caspase-3 免疫印迹测量 5-FU 在 3 种人结肠癌细胞系（HCT116、DLD-1 和 DLD-1/5-FU（特定的 5-FU 耐药亚系））中诱导的细胞凋亡。通过微管相关蛋白 1 轻链 3 (LC3) 免疫荧光和免疫印迹以及 p62 免疫印迹揭示了 5-FU 处理诱导的自噬激活。 3-甲基腺嘌呤 (3-MA) 或靶向 Atg7 的小干扰 RNA (Atg7 siRNA) 对自噬的抑制显着增强了 5-FU 诱导的细胞凋亡。 5-FU和3-MA的这种协同作用在DLD-1异种移植肿瘤模型中得到进一步证实。联合治疗比单独使用 5-FU 治疗更能显着抑制肿瘤生长。总之，自噬被激活作为针对 5-FU 诱导的细胞凋亡的保护机制，其抑制可能是结肠癌辅助化疗的一种有前途的策略。 (C) 2010 Elsevier Ltd. 保留所有权利。
Although 5-fluorouracil (5-FU)-based adjuvant chemotherapy is widely used in the treatment of colorectal cancer, novel therapeutic strategies need to be explored. It has been reported that autophagy is extensively implicated in cancer. However, the function of autophagy is not fully understood. In the present study, apoptosis induced by 5-FU in 3 human colon cancer cell lines (HCT116, DLD-1, and DLD-1/5-FU (a specific 5-FU-resistant sub-line)) was measured using MTT assay, DNA fragmentation assay, Hoechst 33342 staining, and caspase-3 immunoblotting. The autophagy activation induced by 5-FU treatment was revealed by microtubule-associated protein 1 light chain 3 (LC3) immunofluorescence and immunoblotting and p62 immunoblotting. Inhibition of autophagy by 3-methyladenine (3-MA) or small interference RNA targeting Atg7 (Atg7 siRNA) significantly augmented 5-FU-induced apoptosis. This synergistic effect of 5-FU and 3-MA was further confirmed in the DLD-1 xenograft tumour model. Tumour growth was suppressed more significantly with combination treatment than 5-FU treatment alone. In conclusion, autophagy was activated as a protective mechanism against 5-FU-induced apoptosis and its inhibition could be a promising strategy for adjuvant chemotherapy in colon cancer. (C) 2010 Elsevier Ltd. All rights reserved.