Short-chain fatty acids reprogram metabolic profiles with the induction of reactive oxygen species production in human colorectal adenocarcinoma cells

Chongyang Huang; Wenjun Deng; Huan-Zhou Xu; Chen Zhou; Fan Zhang; Junfei Chen; Qinjia Bao; Xin Zhou; Maili Liu; Jing Li; Chaoyang Liu

doi:10.1016/j.csbj.2023.02.022

Short-chain fatty acids reprogram metabolic profiles with the induction of reactive oxygen species production in human colorectal adenocarcinoma cells

Comput Struct Biotechnol J. 2023 Feb 13:21:1606-1620. doi: 10.1016/j.csbj.2023.02.022. eCollection 2023.

Authors

Chongyang Huang¹, Wenjun Deng², Huan-Zhou Xu³, Chen Zhou², Fan Zhang⁴, Junfei Chen¹, Qinjia Bao^{1

5}, Xin Zhou^{1

5

6}, Maili Liu^{1

5

6}, Jing Li^{2

5}, Chaoyang Liu^{1

5

6}

Affiliations

¹ State Key Laboratory of Magnetic Resonance and Atomic and Molecular Physics, Innovation Academy for Precision Measurement Science and Technology, Chinese Academy of Sciences, Wuhan, China.
² Wuhan Botanical Garden, Chinese Academy of Sciences, Wuhan, Hubei, China.
³ Department of Pediatrics, Division of Infectious Diseases, College of Medicine, University of Florida, Gainesville, FL, USA.
⁴ Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, China.
⁵ University of Chinese Academy of Sciences, Beijing, China.
⁶ Optics Valley Laboratory, Hubei 430074, China.

Abstract

Short-chain fatty acids (SCFAs) exhibit anticancer activity in cellular and animal models of colon cancer. Acetate, propionate, and butyrate are the three major SCFAs produced from dietary fiber by gut microbiota fermentation and have beneficial effects on human health. Most previous studies on the antitumor mechanisms of SCFAs have focused on specific metabolites or genes involved in antitumor pathways, such as reactive oxygen species (ROS) biosynthesis. In this study, we performed a systematic and unbiased analysis of the effects of acetate, propionate, and butyrate on ROS levels and metabolic and transcriptomic signatures at physiological concentrations in human colorectal adenocarcinoma cells. We observed significantly elevated levels of ROS in the treated cells. Furthermore, significantly regulated signatures were involved in overlapping pathways at metabolic and transcriptomic levels, including ROS response and metabolism, fatty acid transport and metabolism, glucose response and metabolism, mitochondrial transport and respiratory chain complex, one-carbon metabolism, amino acid transport and metabolism, and glutaminolysis, which are directly or indirectly linked to ROS production. Additionally, metabolic and transcriptomic regulation occurred in a SCFAs types-dependent manner, with an increasing degree from acetate to propionate and then to butyrate. This study provides a comprehensive analysis of how SCFAs induce ROS production and modulate metabolic and transcriptomic levels in colon cancer cells, which is vital for understanding the mechanisms of the effects of SCFAs on antitumor activity in colon cancer.

Keywords: 1H–13C HMBC, 1H–13C Heteronuclear Multiple Bond Correlation Spectroscopy; 1H–13C HSQC, 1H–13C Heteronuclear Single Quantum Coherence Spectroscopy; 1H–1H COSY, 1H–1H Correlation Spectroscopy; 1H–1H TOCSY, 1H–1H Total Correlation Spectroscopy; ADP, Adenosine diphosphate; AMP, Adenosine monophosphate; ATP, Adenosine triphosphate; Ace, Acetate; Ach, Acetylcholine; Ala, Alanine; CRC, Colorectal Cancer; Caco-2, Human Colon Adenocarcinoma; Cho, Choline; CoA, Coenzyme A; Cre, Creatine; DCFH-DA, Dichloro-Dihydro-Fluorescein Diacetate; DEGs, Differentially Expressed Genes; DMEM, Dulbecco's Modified Eagle Medium; DMG, Dimethylglycine; DNA, Deoxyribonucleic Acid; EP, Eppendorf; FA, Formate; FDR, False Discovery Rate; Fru, Fructose; Fum, Fumaric acid; GLS, Glutaminase; GSEA, Gene Set Enrichment Analysis; GSH, Glutathione; Gal-1-P, Galactose-1-phosphate; Glc, Glucose; Gln, Glutamine; Glu, Glutamate; Gly, Glycine; HCT116, Human Colorectal Carcinoma Cell Line; HEK, Human Embryonic Kidney cells; HT29, Human Colorectal Adenocarcinoma Cell Line with Epithelial Morphology; His, Histidine; Ile, Isoleucine; J-Res, J-resolved Spectroscopy; LDH, Lactate Dehydrogenase; Lac, Lactate; Leu, Leucine; Lys, Lysine; MCF-7, Human Breast Cancer Cell Line with Estrogen; MCT, Monocarboxylate Transporters; Met, Methionine; MetS, Metabolic Syndrome; Mitochondrial function; NAD+, Nicotinamide adenine dinucleotide; NAG, N-Acetyl-L-Glutamine; NMR, Nuclear Magnetic Resonance; NMR-based Metabolomics; NOESY, Nuclear Overhauser Effect Spectroscopy; O-PLS-DA, Orthogonal Projection to the Latent Structures Discriminant Analysis; PA, Pantothenate; PC, Phosphocholine; PCA, Principal Component Analysis; PDC, Pyruvate Decarboxylase; PDK, Pyruvate Dehydrogenase Kinase; PKC, Protein Kinase C; PPP, Pentose Phosphate Pathway; Phe, Phenylalanine; Pyr, Pyruvate; RNA, Ribonucleic Acid; ROS, Reactive Oxygen Species; RPKM, Reads per Kilobase of Transcript per Million Reads Mapped; Reactive oxygen species; SCFAs, Short Chain Fatty Acids; SLC, Solute-Carrier Genes; Short-chain fatty acids; Suc, Succinate; T2DM, Type 2 Diabetes; TCA, Tricarboxylic Acid; Tau, Taurine; Thr, Threonine; Transcriptomics; Tyr, Tyrosine; UDP, Uridine 5′-diphosphate; UDP-GLC, UDP Glucose; UDPG, UDP Glucuronate; UDPGs, UDP Glucose and UDP Glucuronate; UMP, Uridine 5′-monophosphate; Val, Valine; WST-1, Water-Soluble Tetrazolium salts; dDNP, dissolution Dynamic Nuclear Polarization; qRT-PCR, Real-Time Quantitative Reverse Transcription Polymerase Chain Reaction; α-KIV, α-Keto-isovalerate; α-KMV, α-keto-β-methyl-valerate.