Fig 1: SOAT1, but not SOAT2, has a more important role in the development of HCC and is negatively correlated with FAO pathway on data mining.A Frequency and types of SOAT1 and SOAT2 DNA alteration in TCGA using cBioPortal. B Copy number variation of SOAT1 or SOAT2 in HCC and normal tissue. ****p < 0.0001; ns, no significance. C mRNA expression pattern of SOAT1 (up) vs. SOAT2 (bottom) in 30 cancer types in the TCGA database. D Paired profiles of SOAT1 vs SOAT2 mRNA expression in tumor tissues from individuals with HCC and TGCT in the TCGA database. RSEM, RNA-Seq by Expectation Maximization. E Kaplan–Meier survival plot of HCC patient based on SOAT1 or SOAT2 expression using TCGA database. SOAT1 cutoff is 10.2503. SOAT2 cutoff is 8.109. Significance was analyzed by Log-rank test. F The frequency of SOAT1 DNA alteration in 19 cancer types from the TCGA database. G Enrichment analysis of the genes altered in the SOAT1 neighborhood in HCC, BP biological processes, MF molecular functions, CC cellular component, KEGG KEGG pathway analysis, NES normal enrichment score.
Fig 2: SOAT1 inhibition upregulates CPT1A expression, which shuttles released FFAs into mitochondria for oxidation in vitro.A–B CCK8 assay showed that SOAT1 knockdown inhibited cell viability in HepG2 and HUH7, (mean ± SEM, n = 3), *p < 0.05. IC50 values of avasimibe (AVA) were about 20 μm. C Western blot analysis of CDK4, CDK6, and cyclinD1 protein with AVA treatment. D–E Fluorescence imaging of LDs stained with BODIPY 493/503 (green) in HUH7 treated with pharmacological (avasimibe, 48 h) or genetic (si-RNA, 72 h) inhibition of SOAT1. Nuclei were stained with DAPI (blue). Scale bar, 50 μm. More than 30 cells were analyzed. Data were quantified using ImageJ software; *p < 0.05; ***p < 0.001, ****p < 0.0001. Significance was determined by unpaired student’s t-test. F The level of total free fatty acids in HCC cells treated with SOAT1 inhibition. (mean ± SEM, n = 3), ns, no significance. G–H Western blot analysis of CPT1A and HMGCL in HepG2 and HUH7 treated with pharmacologic or genetic SOAT1 inhibition. I–K Levels of total acetyl-CoA, ATP, and β-hydroxybutyrate in HCC cells treated with avasimibe (AVA) for 48 h at an indicated concentration (mean ± SEM, n = 3); *p < 0.05; ***p < 0.001, ****p < 0.0001; ns, no significance. L Western blot analysis of P-ACC (S79), ACC, P-AMPK (T172), and AMPK in HCC cells treated with AVA for 48 h at the indicated concentration.
Fig 3: Inhibiting CPT1A converts excessive FFAs into lipid drops by SOAT1.A Western blot analysis of CDK4, CDK6, CyclinD1, ACADL, and ACADM in HCC cells treated with ETO at the indicated concentration. B CCK8 assay showed that etomoxir (ETO) inhibited cell viability in the HepG2 and HUH7 (means ± SEM, n = 3); **p < 0.01, ns, no significance. C The level of total free fatty acids in HCC cells treated with CPT1A inhibition (means ± SEM, n = 3). ns: no significance. D–E Fluorescence imaging of LDs stained with BODIPY 493/503 (green) treated with pharmacological (avasimibe, 48 h) or genetic (si-RNA, 72 h) inhibition of SOAT1 in HUH7. Nuclei were stained with DAPI (blue). Scale bar, 50 μm. More than 30 cells were analyzed. Data were quantified using ImageJ software; ***p < 0.001, ****p < 0.0001, ns: no significance. F Western blot analysis of SOAT1 in HepG2 and HUH7 treated with ETO at the indicated concentration for 48 h. G–I Cellular LDs level in HCC cells treated with ETO plus AVA. More than 30 cells were analyzed.; *p < 0.05, ***p < 0.001, ****p < 0.0001.
Fig 4: HFD improves the expression of SOAT1 and CPT1A in DEN-induced HCC.A–B A high-fat diet exacerbated HCC progression in DEN-injected mice. Representative images of the livers (A) and tumor volumes (B), ND: the normal diet group (n = 8) and HFD: the high-fat diet group (n = 5). (mean ± SEM) **p < 0.01. C Kaplan–Meier survival plot of DEN-injected mice on a high-fat diet (HFD) or normal diet (ND); *p < 0.05. D H&E-stained images of livers using HCC and adjacent non-tumor liver tissue in ND-fed or HFD-fed mice (scale bar, 50 μm). NT and T indicated non-tumor and tumor areas, respectively. E Western blot analysis of SOAT1 and CPT1A in liver cancer tissues from ND-fed or HFD-fed mice. F Representative IHC staining (n = 5 images in total) of SOAT1 or CPT1A in the tissues obtained from HFD-fed mice. NT and T indicate non-tumor and tumor areas, respectively (scale bar, 50 μm). Five separate areas from each tissue were quantified (mean ± SEM); *p < 0.05; **p < 0.01, ***p < 0.001.
Fig 5: Combined avasimibe and etomoxir enhance the efficacy by disrupting lipid homeostasis in HCC in vitro.A–B CCK8 assay showed that combined avasimibe and etomoxir enhanced the efficacy in the HepG2 and HUH7 (means ± SEM, n = 3). Chou and Talalay analyzed the combination indices (CI) of combined AVA and ETO for 48 h in HepG2 and HUH7. The straight line at CI = 1 represents the additive effects of both drugs. Fraction effect (FA): growth inhibition rate. C Representative images of the EdU staining assay in HUH7 cells treated with AVA, ETO, or both. D The colony formation assay of HepG2 and HUH7 cells cultured with 2 μM AVA and/or 25 μM ETO for 14 days. E Western blot analysis of CDK4, CDK6, and Cyclin D1 in HepG2 and HUH7 cells treated with 20 μM AVA and/or 150 μM ETO. F Western blot analysis of CyclinD1, CPT1A, and SOAT1 after transfection with si-SOAT1and/or si-CPT1A in PLC/PCF/5 and HUH7. G–I Levels of total acetyl-CoA, ATP and β-hydroxybutyrate in HCC cells treated with AVA (5 μM) and/or ETO (50 μM) (mean ± SEM, n = 3); *p < 0.05, **p < 0.01, ***p < 0.001; ns: no significance. J Levels of intrahepatic FFAs in HepG2 and HUH7 treated with AVA (5 μM) and/or ETO (50 μM) (mean ± SEM, n = 3); *p < 0.05.
Supplier Page from Novus Biologicals, a Bio-Techne Brand for SOAT1 Antibody - BSA Free
Available conjugates: Available conjugates: UnconjugatedSpecificity: This product is specific for protein corresponding to SOAT1 Uniprot# P35610, Gene ID# 6646. [Important Note:- ACAT1 is a symbol for both acyl-Coenzyme A: cholesterol acyltransferase 1 (SOAT1/ACAT1) and for acetyl-CoA acetyltransferase 1 (ACAT1/ THIL, Gene ID 38, Uniprot P24752) which often cause confusion].Sizes Available: 0.1 ml (also 0.025 ml)