AMPK activator AICAR promotes 5-FU-induced apoptosis in gastric cancer cells.
The aim of the present study was to determine the effect of AICAR, an AMPK activator, on apoptosis in gastric carcinoma cells (SGC-7901) with or without 5-fluorouracil (5-FU). SGC-7901 cells were treated with AICAR (0.2-5 mM, for 24-48 h) with or without 5-FU. Cell viability was determined using MTT assay, while apoptosis were measured through the evaluation of active caspase-3 activity and DNA fragmentation. Real-time PCR was employed to determine the expression of tumor suppressor and multi-drug resistant (mdr1) gene. Cleaved caspase-3 and phosphorylated AMPK (p-AMPK) were measured by Western blot. AICAR significant reduced cellular viability but increased apoptosis in a time- and dose-dependent manner, which is associated with an increase in p-AMPK levels. Importantly, AICAR enhanced the sensitivity to 5-FU-induced reduction of cellular viability and increased apoptosis in SGC-7901 cells. Furthermore, AICAR increased tumor suppressor genes [F-box and WD repeat domain containing 7 (FBXW7), semaphorin III/F (SEMA3F), and p21(Cip1) (p21)] but reduced mdr1 expression. Finally, p-AMPK levels were reduced in 5-FU-resistant gastric cancer cells compared to human immortalized gastric epithelial cell line and 5-FU-sensitive gastric cancer cells. AICAR not only induces apoptosis alone but also enhances pro-apoptotic effect of 5-FU in SGC-7901 cells, which lays an experimental foundation to develop AICAR as a chemotherapeutic sensitizer against gastric cancer.
Department of Surgery, First Affiliated Hospital of Anhui Medical University, Hefei, Anhui, People's Republic of China.|Laboratory of Molecular Biology and Department of Biochemistry, Anhui Medical University, Hefei, Anhui, People's Republic of China.
Yan
Y
Wu
Department of Surgery, First Affiliated Hospital of Anhui Medical University, Hefei, Anhui, People's Republic of China.
Yijun
Y
Qi
Department of Surgery, First Affiliated Hospital of Anhui Medical University, Hefei, Anhui, People's Republic of China.
Hu
H
Liu
Department of Surgery, First Affiliated Hospital of Anhui Medical University, Hefei, Anhui, People's Republic of China.
Xiaoshan
X
Wang
Laboratory of Molecular Biology and Department of Biochemistry, Anhui Medical University, Hefei, Anhui, People's Republic of China. aydzhq@126.com.
Huaqing
H
Zhu
Department of Surgery, First Affiliated Hospital of Anhui Medical University, Hefei, Anhui, People's Republic of China. wangzhengguang@ahmu.edu.cn.
Zhengguang
Z
Wang
D000263:Adenylate Kinase / Q000378:metabolism*; D000620:Aminoimidazole Carboxamide / Q000031:analogs & derivatives* / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects*; D002460:Cell Line; D004789:Enzyme Activation; D005472:Fluorouracil / Q000494:pharmacology*; D006801:Humans; D010766:Phosphorylation; D012265:Ribonucleotides / Q000494:pharmacology*; D013274:Stomach Neoplasms / Q000473:pathology*
D012265:Ribonucleotides; D000620:Aminoimidazole Carboxamide; D000263:Adenylate Kinase; C031143:AICA ribonucleotide; D005472:Fluorouracil
AMPK; Caspase-3; Chemosensitivity; Gastric carcinoma; Multidrug resistance