Department of Gastroenterology, The First Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330006, P.R. China.
Mol Med Rep. 2018 May;17(5):6768-6776. doi: 10.3892/mmr.2018.8661. Epub 2018 Mar 1.
The exact molecular mechanism of 5-fluorouracil (5-FU) in human gastric cancer cells remains to be elucidated. Cultured BGC‑823 human gastric carcinoma and AGS cell lines were treated with 5‑FU. Autophagosome formation was investigated through multiple approaches, including the quantification of green fluorescent protein‑microtubule‑associated protein 1A/1B‑light chain 3 (LC3) puncta, LC3 conversion and electron microscopy observations. Additionally, autophagy was inhibited using 3‑methyladenine (3‑MA) and beclin‑1 ablation, to determine its role in 5‑FU‑mediated cell death. In addition, the present study assessed alterations in sirtuin expression following 5‑FU treatment with reverse transcription‑quantitative polymerase chain reaction. 5‑FU treatment induced apoptosis and inhibited proliferation in BGC‑823 and AGS gastric cancer cells. It is of note that the 5‑FU treatment only promoted autophagy in BGC‑823 cells. Additionally, inhibition of autophagy by either 3‑MA or beclin‑1 ablation increased 5‑FU‑induced cell death in BGC‑823 cells. The present study quantified changes in sirtuin (SIRT1, SIRT3, SIRT5, and SIRT6) expression following 5‑FU treatment and using a specific inhibitor, sirtinol, the present study investigated their involvement in 5‑FU‑mediated autophagy. Autophagy inhibition through manipulation of sirtuin proteins may increase the therapeutic efficacy of the 5‑FU chemotherapeutic drug against gastric carcinoma.
5-氟尿嘧啶（5-FU）在人胃癌细胞中的确切分子机制仍有待阐明。用 5-FU 处理 BGC-823 人胃癌细胞和 AGS 细胞系。通过多种方法研究自噬体的形成，包括绿色荧光蛋白-微管相关蛋白 1A/1B-轻链 3（LC3）斑点的定量、LC3 转化和电子显微镜观察。此外，通过 3-甲基腺嘌呤（3-MA）和 beclin-1 缺失抑制自噬，以确定其在 5-FU 介导的细胞死亡中的作用。此外，本研究还通过逆转录定量聚合酶链反应评估了 5-FU 处理后沉默调节蛋白表达的变化。5-FU 处理诱导 BGC-823 和 AGS 胃癌细胞凋亡和增殖抑制。值得注意的是，5-FU 处理仅在 BGC-823 细胞中促进自噬。此外，通过 3-MA 或 beclin-1 缺失抑制自噬增加了 BGC-823 细胞中 5-FU 诱导的细胞死亡。本研究定量分析了 5-FU 处理后沉默调节蛋白（SIRT1、SIRT3、SIRT5 和 SIRT6）表达的变化，并使用特异性抑制剂 sirtinol 研究了它们在 5-FU 介导的自噬中的作用。通过操纵沉默调节蛋白来抑制自噬可能会增加 5-FU 化疗药物对胃癌的治疗效果。
