PMID- 37272103 OWN - NLM STAT- MEDLINE DCOM- 20230606 LR - 20230612 IS - 1944-7930 (Electronic) IS - 1539-6509 (Linking) VI - 35 IP - 176 DP - 2023 Jun TI - FABP4 Regulates Cell Proliferation, Stemness, Apoptosis, and Glycolysis in Colorectal Cancer via Modulating ROS/ERK/mTOR Pathway. PG - 361-371 LID - 10.24976/Discov.Med.202335176.37 [doi] AB - BACKGROUND: Colorectal cancer is a common digestive tract malignancy. This study aimed to expound the functional role of fatty-acid-binding protein 4 (FABP4) and the potential underlying mechanisms in the development of colorectal cancer. METHODS: Several techniques were utilized to investigate the role of FABP4 in colorectal cancer. FABP4 mRNA expression was quantified using Real time-quantitative PCR (RT-qPCR). Cell counting kit-8 (CCK-8), 5-ethynyl-2'-deoxyuridine (EdU), sphere formation assays and flow cytometry evaluated cell growth, stemness, and apoptosis in SW480 and HT29 cells. Glycolysis was assessed via extracellular acidification rate (ECAR) , lactate production, glucose uptake, adenosine triphosphate (ATP)/adenosine 5'-diphosphate (ADP) ratio, and Glut1 and Elevated lactate dehydrogenase A (LDHA) protein expression. Reactive oxygen species (ROS) levels were analyzed by flow cytometry. Western blot measured the protein expression of FABP4, Proliferating cell nuclear antigen (PCNA), Bax, Bcl-2, Glut1, LDHA, stemness makers (Sox2, Oct4, and ALDHA1), and extracellular regulated protein kinase (ERK)/mammalian target of rapamycin (mTOR) pathway proteins. In vivo experiments, BALB/c nude mice (n = 12) were inoculated with 200 muL HT29 cells (5 x 10(6) cells) transfected with sh-FABP4 or short hairpin (sh)-negative control (NC), forming two groups with 6 mice each. The in vivo mice tumor model allowed for evaluating FABP4's impact on tumor growth. RESULTS: FABP4 was significantly upregulated in colorectal cancer tissues and cells (p < 0.05). FABP4 knockdown markedly inhibited cell proliferation, stemness, and glycolysis, while promoting apoptosis in these cells (p < 0.05). Additionally, FABP4 depletion led to a significant increase in ROS level (p < 0.05). However, N-acetyl-L-cysteine (NAC) (p < 0.05), a ROS scavenger, mitigates these effects. Furthermore, the effects of FABP4 depletion on cell growth, stemness, glycolysis, and apoptosis in colorectal cancer cells were also retarded by NAC (p < 0.05). Notably, FABP4 knockdown also suppressed the ERK/mTOR pathway, suggesting its regulation via ROS (p < 0.05). In vivo study results showed, FABP4 depletion significantly curbed tumor growth in colorectal cancer (p < 0.05). CONCLUSIONS: These results suggest that FABP4 depletion inhibits colorectal cancer progression by modulating cell growth, stemness, glycolysis and apoptosis. This regulation occurs through the ROS/ERK/mTOR pathway. CI - (c) 2023 The Author(s). Published by Discovery Medicine. FAU - Gao, Yingchao AU - Gao Y AD - Department of Gastrointestinal Surgery, Hebei Key Laboratory of Colorectal Cancer Precision Diagnosis and Treatment, The First Hospital of Hebei Medical University, 050031 Shijiazhuang, Hebei, China. FAU - Wang, Yuanyuan AU - Wang Y AD - Department of Gastrointestinal Surgery, Hebei Key Laboratory of Colorectal Cancer Precision Diagnosis and Treatment, The First Hospital of Hebei Medical University, 050031 Shijiazhuang, Hebei, China. FAU - Wang, Xin AU - Wang X AD - Department of Pathology, Hebei Key Laboratory of Colorectal Cancer Precision Diagnosis and Treatment, The First Hospital of Hebei Medical University, 050031 Shijiazhuang, Hebei, China. FAU - Ma, Jianwei AU - Ma J AD - Department of Gastrointestinal Surgery, Hebei Key Laboratory of Colorectal Cancer Precision Diagnosis and Treatment, The First Hospital of Hebei Medical University, 050031 Shijiazhuang, Hebei, China. FAU - Wei, Ming AU - Wei M AD - Department of Gastrointestinal Surgery, Hebei Key Laboratory of Colorectal Cancer Precision Diagnosis and Treatment, The First Hospital of Hebei Medical University, 050031 Shijiazhuang, Hebei, China. FAU - Li, Na AU - Li N AD - Department of Gastrointestinal Surgery, Hebei Key Laboratory of Colorectal Cancer Precision Diagnosis and Treatment, The First Hospital of Hebei Medical University, 050031 Shijiazhuang, Hebei, China. FAU - Zhao, Zengren AU - Zhao Z AD - Department of Gastrointestinal Surgery, Hebei Key Laboratory of Colorectal Cancer Precision Diagnosis and Treatment, The First Hospital of Hebei Medical University, 050031 Shijiazhuang, Hebei, China. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't PL - United States TA - Discov Med JT - Discovery medicine JID - 101250006 RN - 0 (Reactive Oxygen Species) RN - 0 (Glucose Transporter Type 1) RN - EC 2.7.11.1 (TOR Serine-Threonine Kinases) RN - 0 (Fabp4 protein, mouse) RN - 0 (Fatty Acid-Binding Proteins) SB - IM MH - Animals MH - Mice MH - *Signal Transduction MH - Reactive Oxygen Species/metabolism MH - Glucose Transporter Type 1/metabolism MH - Mice, Nude MH - TOR Serine-Threonine Kinases/metabolism MH - Cell Proliferation MH - Apoptosis MH - *Colorectal Neoplasms/genetics/pathology MH - Glycolysis MH - Cell Line, Tumor MH - Mammals/metabolism MH - Fatty Acid-Binding Proteins/genetics/metabolism/pharmacology OTO - NOTNLM OT - ERK/mTOR pathway OT - FABP4 OT - NAC OT - colorectal cancer COIS- The authors declare no conflict of interest. EDAT- 2023/06/05 06:43 MHDA- 2023/06/06 06:42 CRDT- 2023/06/05 03:41 PHST- 2023/06/06 06:42 [medline] PHST- 2023/06/05 06:43 [pubmed] PHST- 2023/06/05 03:41 [entrez] AID - 1685356143376-667277949 [pii] AID - 10.24976/Discov.Med.202335176.37 [doi] PST - ppublish SO - Discov Med. 2023 Jun;35(176):361-371. doi: 10.24976/Discov.Med.202335176.37.