Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Ablation of Sympathetic Nerve-β3 Adrenergic Receptor-mediated Adipose Tissue Lipolysis Attenuates Alcohol-induced Liver Injury in Mice

doi: 10.1016/j.jcmgh.2025.101623

Figure Lengend Snippet: Ablation of Adrb3 in adipose tissues attenuates acute alcohol-induced hepatic steatosis. Chow-fed Adrb3 fl/fl and Adrb3 FKO mice were given a single alcohol binge. ( A ) Liver FATP5 protein expression. ( B ) Liver FFA. ( C ) Liver triglyceride. ( D ) Plasma ALT. ( E ) Plasma triglyceride. ( F ) H&E staining of liver tissue sections. Scale bar: 50 μm. ( G ) Oil Red O staining of liver tissue sections. Scale bar: 200 μm. ( H–K ) RT-qPCR was performed using liver samples for the detection of ( H ) SREBP1c, ACC, FASN, and SCD1; ( I ) DGAT1 and DGAT2; ( J ) PPARα, CPT1α, and Acox1; ( K ) ApoB. n = 4–6. Data are expressed as the means ± SEM. ∗ P < .05, ∗∗ P < .01, ∗∗∗ P < .001.

Article Snippet: Primary antibody FATP5 (#NBP2-37412) was purchased from Novus Biologicals.

Techniques: Expressing, Clinical Proteomics, Staining, Quantitative RT-PCR

Journal: Scientific Reports

Article Title: Intriguing hepatoprotective effects of sucrose on hepatocellular carcinoma pathogenesis

doi: 10.1038/s41598-024-74991-4

Figure Lengend Snippet: Genes related to oxidative stress response and lipid regulation. ( a ) Protein-protein interaction network illustrating key regulators of oxidative stress and lipid metabolism. ( b ) mRNA expression levels: Pgc1a decreased in all groups vs. 18-C, while Pgc1b showed no significant changes. PPRC1 significantly increased during the neoplastic period (18-18-D/S). Sucrose co-administration (18-6-D/S and 18-18-D/S) led to marked differences in Fabp1 and Cav1 expression vs. 18-DEN. ( c ) Disease-free progression in HCC patients: Alterations in genes such as PPARGC1A, PPARGC1B, PPRC1, PPARA, PPARD, PPARG, SLC27A5, FABP1, CAV1, and CD36 correlated with increased disease progression compared to patients without these gene alterations . FC Fold change.

Article Snippet: TaqMan ® gene expression assays were used in the QuantStudio 7 Flex Real-time PCR system (Applied Biosystem, MA, USA) to evaluate the expression of the following rat genes, Ggt1 (Rn00587709_m1), Gstp1 (Rn00561378_gH), Keap1 (Rn00589292_m1), and Nfe2l2 (Rn00477784_m1), Ppargc1a (Rn00580241_m1), Ppargc1b (Rn00598552_m1), Pprc1 (Rn01411002_m1), Ppara (Rn00566193_m1), Ppard (Rn00565707_m1), Pparg (Rn00440945 _m1), Slc27a5 (Rn00577177_m1), Fabp1 (Rn00664587_m1), Cd36 (Rn00580728_m1) and Cav1 (Rn00755834_m1).

Techniques: Expressing, Biomarker Discovery

Journal: Aging (Albany NY)

Article Title: Copper metabolism-related risk score identifies hepatocellular carcinoma subtypes and SLC27A5 as a potential regulator of cuproptosis

doi: 10.18632/aging.205334

Figure Lengend Snippet: The expression and function of SLC27A5 in HCC. ( A – C ) The transcriptional expression of SLC27A5 in HCC and normal tissues of TCGA-LIHC ( A ), GSE14520 ( B ) and GSE64041 ( C ) datasets. ( D ) The protein level of SLC27A5 in HCC and normal tissues. ( E , F ) CCK-8 assay for HepG2 ( E ) and LM-3 ( F ) cells overexpressing SL27A5. ( G , H ) HepG2 ( G ) and LM-3 ( H ) cells were subject to flow cytometry analysis for cell cycle. ( I ) HCC cells overexpressing SLC27A5 were subject to EdU staining. ( J ) HCC cells overexpressing SLC27A5 were subject to wound healing assay.

Article Snippet: Western blotting was performed using antibodies against FDX1 (M05441, Boster), SLC27A5 (A09287-2, Boster), GAPDH (BM3876, Boster).

Techniques: Expressing, CCK-8 Assay, Flow Cytometry, Staining, Wound Healing Assay

Journal: Aging (Albany NY)

Article Title: Copper metabolism-related risk score identifies hepatocellular carcinoma subtypes and SLC27A5 as a potential regulator of cuproptosis

doi: 10.18632/aging.205334

Figure Lengend Snippet: SLC27A5 upregulates FDX1 in HCC. ( A , B ) The correlation between the expression of SLC27A5 and that of FDX1 in TCGA-LIHC ( A ) and GSE14520 ( B ) datasets. ( C ) Dot plot showing the dependency scores for SLC27A5 and FDX1 across all tumor cell lines in the Project Achilles/Cancer Dependency Map Portal (DepMap). ( D , E ) The expression of and correlation between SLC27A5 and FDX1 in collected HCC tissue chip. ( F , G ) HepG2 ( F ) and LM-3 ( G ) cells overexpressing SLC27A5 were subject to qRT-PCR analysis. ( H , I ) Western blotting using lysates of HepG2 ( H ) and LM-3 ( I ) cells after overexpressing SLC27A5.

Article Snippet: Western blotting was performed using antibodies against FDX1 (M05441, Boster), SLC27A5 (A09287-2, Boster), GAPDH (BM3876, Boster).

Techniques: Expressing, Quantitative RT-PCR, Western Blot

Journal: Cell Death and Differentiation

Article Title: SLC27A5 deficiency activates NRF2/TXNRD1 pathway by increased lipid peroxidation in HCC

doi: 10.1038/s41418-019-0399-1

Figure Lengend Snippet: SLC27A5 expression is generally downregulated in human HCC tissues and correlates with poor patient prognosis. a Expression of SLC27A5 in patients with HCC based on data from TCGA Liver Hepatocellular Carcinoma dataset. RKPM, Reads per kilobase of exon model per million mapped reads. ***P < 0.001, two-tailed t-test. b Box plots of SLC27A5 expression in normal liver and tumor tissues (Clinical stages I to III) from the TCGA Database. Thirteen patients with stage IV cancer were excluded from the analysis. c SLC27A5 mRNA expression in 40 paired HCC (T) and nontumorous (NT) liver tissues. ***P < 0.001, Wilcoxon’s signed-rank test. d IHC staining of SLC27A5 in HCC tissues and adjacent nontumor tissues, bar = 50 µm. Immunostaining intensity was assessed using Image-Pro plus 6.0 software. ***P < 0.001, Student’s t-test. e SLC27A5 expression levels in tumor samples were inversely correlated with the degree of SLC27A5 methylation in TCGA (Pearson correlation coefficient = −0.47; P < 0.001). f Nucleotide sequence of SLC27A5 that forms a part of CpG islands around the promoter region. Individual CpG loci are highlighted in red. g Direct bisulfite sequencing of SLC27A5 promoter region for individual CpG loci in HCC tissues (T) and adjacent nontumor tissues (NT). MI: methylation index = (number of methylated CpG loci)/(total number of CpG loci). h Kaplan–Meier survival analysis of TCGA cohort; HCC patients were divided into two groups on the basis of the SLC27A5 expression level in tumor (top 25%, high; bottom 25%, low)

Article Snippet: The full-length cDNA of SLC27A5 (NM_012254.2) was amplified from plasmid EX-W0248-M02 (GeneCopoeia, Guangzhou, China) and inserted into the Kpn I and Hind III sites of the shuttle vector pAdTrack-TO4 (from Dr T-C He, University of Chicago, USA).

Techniques: Expressing, Two Tailed Test, Immunohistochemistry, Immunostaining, Software, Methylation, Sequencing, Methylation Sequencing

Journal: Cell Death and Differentiation

Article Title: SLC27A5 deficiency activates NRF2/TXNRD1 pathway by increased lipid peroxidation in HCC

doi: 10.1038/s41418-019-0399-1

Figure Lengend Snippet: Clinical correlation between mRNA levels of SLC27A5 and clinicopathologic parameters of HCC patients

Article Snippet: The full-length cDNA of SLC27A5 (NM_012254.2) was amplified from plasmid EX-W0248-M02 (GeneCopoeia, Guangzhou, China) and inserted into the Kpn I and Hind III sites of the shuttle vector pAdTrack-TO4 (from Dr T-C He, University of Chicago, USA).

Techniques: Expressing

Journal: Cell Death and Differentiation

Article Title: SLC27A5 deficiency activates NRF2/TXNRD1 pathway by increased lipid peroxidation in HCC

doi: 10.1038/s41418-019-0399-1

Figure Lengend Snippet: SLC27A5 suppresses hepatoma cell proliferation in vitro and in vivo. Ectopic expression of SLC27A5 inhibits cell proliferation (a) and colony formation (b) in PLC/PRF/5 and SK-Hep1 cells (n = 3). SLC27A5 knockout promotes cell proliferation (c) and colony formation (d) (n = 3). MHCC-97H cells were infected with AdSLC27A5 or AdGFP for 36 h. Cells (5 × 106 per site) were subcutaneously injected into the flanks of nude mice (n = 6 per group). e Representative tumor-bearing mice of the subcutaneous tumor model treated as indicated. f, g Graphs (mean ± SD) showing tumor growth and tumor weight. n = 6. h Ki67 staining in tumor tissues, bar = 50 µm. i Orthotopic liver implantations established from SLC27A5-KO HepG2 cells. Representative liver tumors derived from SLC27A5-KO or parental HepG2 cells. j Graphs (mean ± SD) showing liver tumor nodules, n = 6. k Ki67 staining of liver tumor tissues, bar = 50 µm. Data are means ± SDs. P values were derived from repeated-measures ANOVA in a, c, and f; one-way ANOVA test in b, c, d, and g; unpaired, two-tailed Student’s t-test in j; (*P < 0.05, **P < 0.01, ***P < 0.001)

Article Snippet: The full-length cDNA of SLC27A5 (NM_012254.2) was amplified from plasmid EX-W0248-M02 (GeneCopoeia, Guangzhou, China) and inserted into the Kpn I and Hind III sites of the shuttle vector pAdTrack-TO4 (from Dr T-C He, University of Chicago, USA).

Techniques: In Vitro, In Vivo, Expressing, Knock-Out, Infection, Injection, Staining, Derivative Assay, Two Tailed Test

Journal: Cell Death and Differentiation

Article Title: SLC27A5 deficiency activates NRF2/TXNRD1 pathway by increased lipid peroxidation in HCC

doi: 10.1038/s41418-019-0399-1

Figure Lengend Snippet: Loss of SLC27A5 enriches polyunsaturated lipids and elevates oxidative stress and lipid peroxidation. FAO levels in SLC27A5-OE PLC/PRF/5 (a) and SLC27A5-knockout HepG2 cells (b) after nutrient starvation. Increases in maximal respiration beyond the BSA control indicate utilization of exogenous FAs (n = 3). Heatmap of lipid abundances (c) and ceramide (d) in SLC27A5-KO and parental HepG2 cells (n = 6). DG diacylglyceride, So sphingosine, TG triacylglyceride, PA phosphatidic acids, PC phosphatidylcholine, ChE cholesteryl esters, PE phosphatidylethanolamine, PI phosphatidylinositol, PS phosphatidylserine, CL cardiolipins, PG phosphatidylglycerols, Cer ceramides, SM sphingomyelin, SFA saturated fatty acid, MUFA monounsaturated fatty acid, PUFA polyunsaturated fatty acid. e–h Levels of NADP+/NADPH, ROS, and lipid peroxidation in SLC27A5-OE cells. e NADP+/NADPH ratio in SLC27A5-OE cells. f Intracellular ROS were measured by CellROX Orange fluorogenic probe. Scale bar: 10 µM. Mean fluorescence intensity was analyzed with Image Pro Plus software. g MDA levels in SLC27A5-OE cells. MDA concentrations were normalized to total protein content (n = 3, performed in triplicate). h 4-HNE-induced protein modification was examined by western blot analysis. Levels of NADP+/NADPH ratio (i), ROS (j), and lipid peroxidation, including MDA levels (k) and 4-HNE-induced protein modification (l), in SLC27A5-knockout HepG2 cells (n = 3). Data are presented as means ± SDs. P values were derived from an unpaired, two-tailed Student’s t-test in a, b, e, and i; one-way ANOVA test were performed in f, g, j and k (*P < 0.05, ***P < 0.001)

Article Snippet: The full-length cDNA of SLC27A5 (NM_012254.2) was amplified from plasmid EX-W0248-M02 (GeneCopoeia, Guangzhou, China) and inserted into the Kpn I and Hind III sites of the shuttle vector pAdTrack-TO4 (from Dr T-C He, University of Chicago, USA).

Techniques: Knock-Out, Fluorescence, Software, Modification, Western Blot, Derivative Assay, Two Tailed Test

Journal: Cell Death and Differentiation

Article Title: SLC27A5 deficiency activates NRF2/TXNRD1 pathway by increased lipid peroxidation in HCC

doi: 10.1038/s41418-019-0399-1

Figure Lengend Snippet: SLC27A5 inhibits NRF2 antioxidative pathway. a, b NRF2-mediated gene transcriptional activities in SLC27A5-OE or -KO HCC cells as assessed by ARE-luciferase assays (n = 3). Tert-butylhydroquinone (tBHQ, 100 µM) was used as a positive control. c Intracellular localization of NRF2 (red) in SLC27A5-OE PLC/PRF/5 cells by the immunofluorescence assay. Nuclei are stained blue with DAPI. Bar = 50 μm. d Intracellular localization of NRF2 (red) in SLC27A5-KO HepG2 cells. Nuclei are stained blue with DAPI. Bar = 10 μm. e, f mRNA expression of NRF2-downstream target genes NQO1, HO-1, GCLM, Me1, TXNRD1, G6PD, and MRP2 in SLC27A5-OE PLC/PRF/5 (e) and SLC27A5-KO HepG2 cells (f). (n = 3, performed in triplicate). Protein expression of NRF2-downstream target genes NQO1, HO-1, and TXNRD1 in SLC27A5-OE PLC/PRF/5 (g) and SLC27A5-KO HepG2 cells (h). i Representative IHC staining of SLC27A5, TXNRD1, and NQO1 in serial tissue slices. HCC tissues (T) and adjacent nontumor tissues (N) (bar = 50 µm). Data are means ± SDs. P values were derived from one-way ANOVA test in a, b, and f; an unpaired, two-tailed Student’s t-test in e (*P < 0.05, **P < 0.01, ***P < 0.001)

Article Snippet: The full-length cDNA of SLC27A5 (NM_012254.2) was amplified from plasmid EX-W0248-M02 (GeneCopoeia, Guangzhou, China) and inserted into the Kpn I and Hind III sites of the shuttle vector pAdTrack-TO4 (from Dr T-C He, University of Chicago, USA).

Techniques: Luciferase, Positive Control, Immunofluorescence, Staining, Expressing, Immunohistochemistry, Derivative Assay, Two Tailed Test

Journal: Cell Death and Differentiation

Article Title: SLC27A5 deficiency activates NRF2/TXNRD1 pathway by increased lipid peroxidation in HCC

doi: 10.1038/s41418-019-0399-1

Figure Lengend Snippet: SLC27A5 represses oxidative modification of KEAP1 by 4-HNE in hepatoma cells. Immunoprecipitation of 4-HNE-modified KEAP1. SLC27A5-overexpressing PLC/PRF/5 (a) and SLC27A5-knockout HepG2 cells (b) were infected with adenoviruses expressing Flag-KEAP1 for 48 h, followed by Flag immunoprecipitation and western blot analysis of 4-HNE. c 4-HNE alkylation of human KEAP1 cysteine residues Cys513 and Cys518 was identified in SLC27A5-knockout HepG2 cells by MS/MS analysis. d Schematic diagram showing the NTR, BTB, IVR, Kelch, and CTR domains of KEAP1. The cysteine residues as well as KEAP1 mutants with substitution of a cysteine (C) to alanine (A) or cysteine (C) to serine (S) are indicated. e, f Constructs expressing Flag-KEAP1 (WT), or Flag-KEAP1 C151A, C151S, C513A, C513S, C518A, or C518S mutant were cotransfected into PLC/PRF/5 cells with plasmids expressing Myc-Nrf2. At 48 h after transfection, cells were treated with 4-HNE (20 µM, 2 h) before blotting for Myc-Nrf2

Article Snippet: The full-length cDNA of SLC27A5 (NM_012254.2) was amplified from plasmid EX-W0248-M02 (GeneCopoeia, Guangzhou, China) and inserted into the Kpn I and Hind III sites of the shuttle vector pAdTrack-TO4 (from Dr T-C He, University of Chicago, USA).

Techniques: Modification, Immunoprecipitation, Knock-Out, Infection, Expressing, Western Blot, Tandem Mass Spectroscopy, Construct, Mutagenesis, Transfection

Journal: Cell Death and Differentiation

Article Title: SLC27A5 deficiency activates NRF2/TXNRD1 pathway by increased lipid peroxidation in HCC

doi: 10.1038/s41418-019-0399-1

Figure Lengend Snippet: SLC27A5 negatively correlates with TXNRD1 expression in hepatoma cells and HCC tissues. mRNA and protein expression of TXNRD1 in SLC27A5-overexpressing cells (a, b) and SLC27A5-KO cells (c, d). Data represent the means ± SD. (n = 3, performed in triplicate). *P < 0.05, ***P < 0.001. e Immunoblot analysis of TXNRD1 in mouse embryonic fibroblasts (MEFs) lysates from Nrf2+/+ and Nrf2−/− cells infected with AdSLC27A5 or AdGFP. β-actin was used as an internal control. f The mRNA expression of SLC27A5 and TXNRD1 in 40 pairs of HCC tissues and adjacent nontumorous tissues (HCC/NT). Data are means ± SD. ***P < 0.001, Wilcoxon’s signed-rank tests were used. g Correlation analysis of SLC27A5 and TXNRD1 mRNA expression in 40 paired HCC and nontumor tissues (*P < 0.05, r = −0.35, Pearson’s correlation). h Immunoblot analysis of SLC27A5 and TXNRD1 protein expression in 20 paired HCC and nontumor tissues. i Representative IHC staining of SLC27A5 and TXNRD1 in serial tissue slices from 20 paired HCC tissues and adjacent nontumor tissues (T/N). (bar = 50 µm)

Article Snippet: The full-length cDNA of SLC27A5 (NM_012254.2) was amplified from plasmid EX-W0248-M02 (GeneCopoeia, Guangzhou, China) and inserted into the Kpn I and Hind III sites of the shuttle vector pAdTrack-TO4 (from Dr T-C He, University of Chicago, USA).

Techniques: Expressing, Western Blot, Infection, Immunohistochemistry

Journal: Cell Death and Differentiation

Article Title: SLC27A5 deficiency activates NRF2/TXNRD1 pathway by increased lipid peroxidation in HCC

doi: 10.1038/s41418-019-0399-1

Figure Lengend Snippet: TXNRD1 knockdown sensitizes hepatoma cells to sorafenib and potentiates apoptosis in vitro and in vivo. TXNRD1 was knocked down in SLC27A5-knockout HepG2 cells using lentiviral shRNA. a, b Sorafenib (Sora) combined with shTXN significantly inhibited cell proliferation (n = 3). c Representative images of orthotopic liver implantation of SLC27A5-knockout HepG2 cells with TXNRD1 knockdown (shTXN) in nude mice. d Liver tumor numbers were assessed (n = 6). e Ki67 staining in liver tumor tissues (n = 3), bar = 50 µm. f TUNEL staining of apoptotic cells in tumor tissues (n = 3), bar = 50 µm. g Representative fluorescence microscopic images of ROS staining in tumor tissues (n = 3). Scale bar: 10 µM. h Proposed working model of this study. LPO lipid peroxidation. All data are means ± SDs. P values were derived from repeated-measures ANOVA in a; one-way ANOVA test in b, d, and g (*P < 0.05, **P < 0.01, ***P < 0.001)

Article Snippet: The full-length cDNA of SLC27A5 (NM_012254.2) was amplified from plasmid EX-W0248-M02 (GeneCopoeia, Guangzhou, China) and inserted into the Kpn I and Hind III sites of the shuttle vector pAdTrack-TO4 (from Dr T-C He, University of Chicago, USA).

Techniques: In Vitro, In Vivo, Knock-Out, shRNA, Staining, TUNEL Assay, Fluorescence, Derivative Assay