hat1 inhibitor jg 2016  (MedChemExpress)

Journal: Oncology Letters

Article Title: Histone acetyltransferase 1 promotes ovarian cancer progression by regulating cell proliferation and the cell cycle

doi: 10.3892/ol.2025.15285

Figure Lengend Snippet: HAT1 levels are upregulated in human ovarian cancer tissue and higher HAT1 levels indicate a poor prognosis. (A) Representative images depicting the representative immunostaining of HAT1 in ovarian cancer tissues and healthy human tissues in The Human Protein Altas database. Scale bar=200 µm. (B) Analysis of the TNMplot database showed that HAT1 was highly expressed in ovarian cancer tissues compared with healthy controls. (C) Protein levels of HAT1 in ovarian cancer and healthy human samples from healthy controls were analyzed in the CPTAC database. (D) HAT1 protein levels were upregulated with tumor grade in ovarian cancer tissues. (E) Higher HAT1 levels were associated with an unfavorable prognosis in patients with ovarian cancer. Data are presented as the mean ± SD. **P<0.01. HAT1, histone acetyltransferase 1; CPTAC, Clinical Proteomic Tumor Analysis Consortium; HR, hazard ratio.

Article Snippet: HEY and SKOV3 cells were seeded into 96-well plates (5×10 3 /well) and treated with 0.0, 0.5, 1.0, 5, 10, 20, 30 and 50 μM) of HAT1 inhibitor JG-2016 or DMSO as indicated (Cat.HY-154944; MedChemExpress) for 72 h at room temperature.

Techniques: Immunostaining

Journal: Oncology Letters

Article Title: Histone acetyltransferase 1 promotes ovarian cancer progression by regulating cell proliferation and the cell cycle

doi: 10.3892/ol.2025.15285

Figure Lengend Snippet: HAT1 expression levels are upregulated in human ovarian cancer cell lines. (A) Relative expression levels of HAT1 were detected by reverse transcription-quantitative PCR in A2780, HEY, OVCAR3, SKOV3 and normal IOSE386 cell lines. (B) Western blot analysis of HAT1 expression in A2780, HEY, OVCAR3, SKOV3 and IOSE386 cell lines. ****P<0.0001 vs. IOSE386. HAT1, histone acetyltransferase 1.

Article Snippet: HEY and SKOV3 cells were seeded into 96-well plates (5×10 3 /well) and treated with 0.0, 0.5, 1.0, 5, 10, 20, 30 and 50 μM) of HAT1 inhibitor JG-2016 or DMSO as indicated (Cat.HY-154944; MedChemExpress) for 72 h at room temperature.

Techniques: Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot

Journal: Oncology Letters

Article Title: Histone acetyltransferase 1 promotes ovarian cancer progression by regulating cell proliferation and the cell cycle

doi: 10.3892/ol.2025.15285

Figure Lengend Snippet: FOXA1 transcriptionally regulates HAT1 expression levels. (A) FOXA1 binding sites in the HAT1 promoter region were predicted using the JASPAR database. MUT constructs were generated at the binding sequence regions as indicated. (B) The expression levels of FOXA1 in HEY cells transfected with an empty or FOXA1 vector. (C) HEY cells were transfected with pmirGLO reporter vectors containing either WT or MUT plasmids alongside an empty or FOXA1 vector. Luciferase activities were determined 24 h after transfection. (D) Overexpression of FOXA1 induced HAT1 expression levels. (E) cBioPortal database was adopted to analyze the correlation between FOXA1 and HAT1. Pearson's rank correlation between HAT1 and FOXA1 was analyzed in ovarian cancer tissues. (F) FOXA1 levels in ovarian cancer tissues from TCGA were determined using TNMplot database. (G) FOXA1 was enriched at the HAT1 promoter region as suggested by Cistrome DB. Data are presented as the mean ± SD of three replicates. **P<0.01 vs. vector or as indicated; ns, not significant. ChIP-seq, chromatin immunoprecipitation sequencing; FOXA1, forkhead box protein A1; HAT1, histone acetyltransferase 1; WT, wild-type; MUT, mutant.

Article Snippet: HEY and SKOV3 cells were seeded into 96-well plates (5×10 3 /well) and treated with 0.0, 0.5, 1.0, 5, 10, 20, 30 and 50 μM) of HAT1 inhibitor JG-2016 or DMSO as indicated (Cat.HY-154944; MedChemExpress) for 72 h at room temperature.

Techniques: Expressing, Binding Assay, Construct, Generated, Sequencing, Transfection, Plasmid Preparation, Luciferase, Over Expression, ChIP-sequencing, Mutagenesis

Journal: Oncology Letters

Article Title: Histone acetyltransferase 1 promotes ovarian cancer progression by regulating cell proliferation and the cell cycle

doi: 10.3892/ol.2025.15285

Figure Lengend Snippet: Relationship between HAT1 and DNA replication and pyrimidine metabolism pathways. (A) LinkedOmics database suggested that differentially expressed genes of HAT1 were enriched in several biological processes, such as ‘DNA replication’, ‘pyrimidine metabolism’ and ‘RNA transport’. (B) Gene set enrichment analysis revealed that genes altered by HAT1 were positively associated with ‘DNA replication’ as well as ‘pyrimidine metabolism’ pathways. (C) cBioPortal database was used to analyze the correlations between HAT1 and regulatory proteins of the DNA replication pathway. Pearson's rank correlation between HAT1 and DNA replication-related proteins (PCNA, RPA1 and POLA1) was analyzed in ovarian cancer tissues. (D) Pearson's rank correlation between HAT1 and pyrimidine metabolism-related proteins (TK1, RRM1 and RRM2) was analyzed in ovarian cancer tissues as suggested in the cBioPortal database. HAT1, histone acetyltransferase 1; PCNA, proliferating cell nuclear antigen; RPA1, replication protein A1; POLA1, DNA polymerase α catalytic subunit; TK1, thymidine kinase 1; RRM, ribonucleoside-diphosphate reductase subunit M; NES, normalized enrichment score; FDR, false discovery rate.

Article Snippet: HEY and SKOV3 cells were seeded into 96-well plates (5×10 3 /well) and treated with 0.0, 0.5, 1.0, 5, 10, 20, 30 and 50 μM) of HAT1 inhibitor JG-2016 or DMSO as indicated (Cat.HY-154944; MedChemExpress) for 72 h at room temperature.

Techniques:

Journal: Oncology Letters

Article Title: Histone acetyltransferase 1 promotes ovarian cancer progression by regulating cell proliferation and the cell cycle

doi: 10.3892/ol.2025.15285

Figure Lengend Snippet: Inhibition of HAT1 suppresses cell viability and colony formation in vitro . (A) Western blot analysis of HAT1 expression in HEY and SKOV3 cells transfected with HAT1 siRNA or NC siRNA. (B) Cell Counting Kit-8 assays were performed to determine cell viability after HAT1 was knocked down in HEY and SKOV3 cells. (C) HEY and SKOV3 cells were treated with HAT1 inhibitor JG-2016 for 72 h, which significantly inhibited cell viability. (D) Knockdown of HAT1 decreased the colony formation capacity of HEY and SKOV3 cells. Data are presented as the mean ± SD of three replicates. *P<0.05 and **P<0.01 vs. siNC in figure 5B and DMSO group in figure 5C. HAT1, histone acetyltransferase 1; si, small interfering; NC, negative control.

Article Snippet: HEY and SKOV3 cells were seeded into 96-well plates (5×10 3 /well) and treated with 0.0, 0.5, 1.0, 5, 10, 20, 30 and 50 μM) of HAT1 inhibitor JG-2016 or DMSO as indicated (Cat.HY-154944; MedChemExpress) for 72 h at room temperature.

Techniques: Inhibition, In Vitro, Western Blot, Expressing, Transfection, Cell Counting, Knockdown, Negative Control

Journal: Oncology Letters

Article Title: Histone acetyltransferase 1 promotes ovarian cancer progression by regulating cell proliferation and the cell cycle

doi: 10.3892/ol.2025.15285

Figure Lengend Snippet: HAT1 knockdown inhibits cell proliferation. (A) EdU assays of HEY cells were performed showing that suppression of HAT1 attenuated cell proliferation activities. Scale bar, 20 µm. (B) Cell cycle analysis was carried out on the HAT1 knockdown and siNC cell lines, and the distribution of G 0 /G 1 , S and G 2 /M percentages were analyzed. (C) Western blot analysis of cell cycle-related protein expression after knockdown of HAT1. (D) Pearson's rank correlation between HAT1 and CDK2, CDK4 and cyclin E was analyzed in ovarian cancer tissues. Data are presented as the mean ± SD of three replicates. **P<0.01 vs. siNC. HAT1, histone acetyltransferase 1; si, small interfering; NC, negative control; CDK, cyclin-dependent kinase.

Article Snippet: HEY and SKOV3 cells were seeded into 96-well plates (5×10 3 /well) and treated with 0.0, 0.5, 1.0, 5, 10, 20, 30 and 50 μM) of HAT1 inhibitor JG-2016 or DMSO as indicated (Cat.HY-154944; MedChemExpress) for 72 h at room temperature.

Techniques: Knockdown, Cell Cycle Assay, Western Blot, Expressing, Negative Control

Journal: Acta Pharmaceutica Sinica. B

Article Title: Succinylation of tumor suppressor PPP2R1A K541 by HAT1 converses the role in modulation of gluconeogenesis/lipogenesis remodeling to display oncogene function

doi: 10.1016/j.apsb.2025.07.040

Figure Lengend Snippet: HAT1 is essential for DEN/CCl 4 -induced hepatocarcinogenesis. (A) The scheme depicting the experimental outline of the DEN/CCl 4 -induced HCC mouse model was shown. The wild type (WT) and liver-specific Hat1 knockout C57BL/6 mice ( Hat1 −/− ) mice were administered DEN (25 mg/kg) by i.p. injection on Day 14 after birth and then were administered CCl 4 (0.5 mL/kg) by intraperitoneal injection for 20 weeks after administration of DEN. The mice were analyzed at 48 weeks. (B) The expression of HAT1 was examined by Western blot analysis in liver tissues of WT ( n = 6) and Hat1 −/− mice ( n = 6) from DEN/CCl 4 -induced HCC model. (C) Representative PET/CT images of WT and Hat1 −/− mice from the DEN/CCl 4 -induced HCC mouse model were shown, and the SUV max values were analyzed in the mice. (D) Representative livers from WT and Hat1 −/− mice of DEN/CCl4-induced HCC model were shown. H&E staining and IHC staining of HAT1 and Ki-67 were determined in liver tissues from the mice. Scale bar, 50 μm. (E) The total tumor number and the tumor number (>3 mm) were calculated in the liver of WT ( n = 6) and Hat1 −/− mice from the DEN/CCl 4 -induced HCC model ( n = 6). Each point is one mouse. (F) The liver/body weight ratio was tested in WT and Hat1 −/− mice from the DEN/CCl 4 -induced HCC model. (G, H) Untargeted metabolomics were performed in liver tissues from WT and Hat1 −/− mice of DEN/CCl 4 -induced HCC model. (G) Volcano plot depicting the different levels of metabolites (fold change >1.5, or fold change <0.67, P < 0.05). (H) Enriched Kyoto Encyclopaedia of Genes and Genomes (KEGG) pathways of differently expressed metabolites were presented. Data are presented as mean ± SD. Student's t -test, ∗∗∗ P < 0.001. HAT1: histone acetyltransferase 1, DEN: diethylnitrosamine, CCl 4 : carbon tetrachloride, HCC: hepatocellular carcinoma, WT: wild type, H&E: hematoxylin and eosin (H&E), IHC: immunohistochemistry, KEGG: Kyoto Encyclopaedia of Genes and Genomes.

Article Snippet: Histone acetyltransferase 1 (HAT1) inhibitor JG-2016 was purchased from MCE (Shanghai, China).

Techniques: Knock-Out, Injection, Expressing, Western Blot, Positron Emission Tomography-Computed Tomography, Staining, Immunohistochemistry

Journal: Acta Pharmaceutica Sinica. B

Article Title: Succinylation of tumor suppressor PPP2R1A K541 by HAT1 converses the role in modulation of gluconeogenesis/lipogenesis remodeling to display oncogene function

doi: 10.1016/j.apsb.2025.07.040

Figure Lengend Snippet: HAT1 confers the remodeling of gluconeogenesis/lipogenesis. (A) A model of HAT1 conferring PCK1-mediated remodeling of gluconeogenesis/lipogenesis. (B) The levels of PCK1 pS90, PCK1, HAT1, and β -actin were detected by Western blot analysis in the liver tissues from C57BL/6 mice treated with normal vehicle ( n = 6) or C57BL/6 mice of DEN/CCl 4 -induced HCC model ( n = 6). (C) The levels of PCK1 pS90, PCK1, HAT1, and β -actin were measured by Western blot analysis in liver tissues of WT ( n = 6) and Hat1 −/− mice ( n = 6) from DEN/CCl 4 -induced HCC model. (D) The levels of PCK1 pS90, PCK1, HAT1, and β -actin were determined by Western blot analysis in primary human hepatocytes (PHH), HepG2, and Huh7 cells. (E) The levels of PCK1 pS90, PCK1, HAT1, and β -actin were examined by Western blot analysis in WT HepG2 cells and HAT1 KO Clone 1 HepG2 cells (HAT1 KO #1). (F) The relative levels of PEP, citrate, triglyceride, and cholesterol were analyzed by the corresponding assay kits in the liver tissues of WT ( n = 6) and Hat1 −/− mice ( n = 6) from DEN/CCl 4 -induced HCC model, respectively. (G–J) The relative levels of PEP, citrate, triglyceride, and cholesterol were tested by the corresponding assay kits in WT HepG2 cells and HAT1 KO #1 HepG2 cells cultured in 25 or 0.1 mmol/L glucose conditions for 10 h, respectively. (K) The lipid accumulation was analyzed by Nile Red staining in WT HepG2 cells and HAT1 KO #1 HepG2 cells cultured in 25 or 0.1 mmol/L glucose conditions for 10 h, respectively. Scale bar, 5 μm. The relative Nile Red staining intensity was quantified. Data are presented as mean ± SD. Student's t -test, ∗∗ P < 0.01, ∗∗∗ P < 0.001. HAT1: histone acetyltransferase 1, PCK1: phosphoenolpyruvate carboxykinase 1, pS90: phosphorylation at serine 90, DEN: diethylnitrosamine, CCl 4 : carbon tetrachloride, HCC: hepatocellular carcinoma, WT: wild type, KO: knockout, PEP: phosphoenolpyruvate.

Article Snippet: Histone acetyltransferase 1 (HAT1) inhibitor JG-2016 was purchased from MCE (Shanghai, China).

Techniques: Western Blot, Cell Culture, Staining, Phospho-proteomics, Knock-Out

Journal: Acta Pharmaceutica Sinica. B

Article Title: Succinylation of tumor suppressor PPP2R1A K541 by HAT1 converses the role in modulation of gluconeogenesis/lipogenesis remodeling to display oncogene function

doi: 10.1016/j.apsb.2025.07.040

Figure Lengend Snippet: HAT1 triggers the remodeling of gluconeogenesis/lipogenesis by modulating the levels of phosphorylation of PCK1 Ser90. (A–H) HepG2 cells were transfected with plasmids overexpressing HAT1, or co-transfected with plasmids overexpressing HAT1 and PCK1, PCK1 mutant (S90D), or PCK1 mutant (S90A), respectively. (A) The relative PCK1 activity was analyzed by the corresponding assay kit in the cells. (B) The nuclear accumulation of SREBP1 was observed by confocal immunofluorescence analysis in the cells. Scale bar, 5 μm. (C) The mRNA expression levels of SREBP1 target genes, including FASN , SCD , ACACA , and GPAM , were analyzed by RT-qPCR in the cells. (D–G) The relative levels of PEP, citrate, triglyceride, and cholesterol were measured by the corresponding assay kits in the cells cultured in 25 or 0.1 mmol/L glucose conditions for 10 h, respectively. (H) The lipid accumulation was showed by Nile Red staining in the cells cultured in 25 or 0.1 mmol/L glucose conditions for 10 h, respectively. Scale bar, 5 μm. The relative Nile Red staining intensity was quantified. Data are presented as mean ± SD. Student's t -test, ∗∗ P < 0.01, ∗∗∗ P < 0.001. HAT1: histone acetyltransferase 1, PCK1: phosphoenolpyruvate carboxykinase 1, pS90: phosphorylation at serine 90, SREBP1: sterol regulatory element-binding protein 1, FASN: fatty acid synthase, SCD: stearoyl-CoA desaturase, ACACA: acetyl-CoA carboxylase alpha, GPAM: glycerol-3-phosphate acyltransferase 1.

Article Snippet: Histone acetyltransferase 1 (HAT1) inhibitor JG-2016 was purchased from MCE (Shanghai, China).

Techniques: Phospho-proteomics, Transfection, Mutagenesis, Activity Assay, Immunofluorescence, Expressing, Quantitative RT-PCR, Cell Culture, Staining, Binding Assay

Journal: Acta Pharmaceutica Sinica. B

Article Title: Succinylation of tumor suppressor PPP2R1A K541 by HAT1 converses the role in modulation of gluconeogenesis/lipogenesis remodeling to display oncogene function

doi: 10.1016/j.apsb.2025.07.040

Figure Lengend Snippet: HAT1 succinylates PPP2R1A K541, leading to phosphorylation of PCK1 Ser90. (A) The levels of PPP2R1A succinylation and HAT1 were examined by immunoprecipitation and Western blot analysis as indicated in the liver tissues from C57BL/6 mice treated with normal vehicle ( n = 6) or tumor tissues from C57BL/6 mice of DEN/CCl 4 -induced HCC model ( n = 6). (B) The levels of PPP2R1A succinylation and HAT1 were measured by immunoprecipitation and Western blot analysis as indicated in tumor tissues of WT ( n = 6) and Hat1 −/− mice ( n = 6) from DEN/CCl 4 -induced HCC model, respectively. (C) The PPP2R1A succinylation levels and HAT1 were determined by immunoprecipitation and Western blot analysis as indicated in HepG2 cells (WT), HAT1 KO #1 HepG2 cells, HAT1 KO #1 HepG2 cells reconstituted with wild-type HAT1 or HAT1 mutant (T188A). (D) The levels of PPP2R1A K541 succinylation, PPP2R1A, and HAT1 were measured by Western blot analysis in HepG2 cells (WT), HAT1 KO #1 HepG2 cells, HAT1 KO #1 HepG2 cells reconstituted with wild-type HAT1 or HAT1 mutant (T188A). (E) HAT1-catalyzed PPP2R1A succinylation was analyzed by in vitro succinylation assays using purified HAT1, PPP2R1A, and succinyl-CoA/acetyl-CoA as indicated. Western blot analysis was performed using the indicated antibodies. (F) The interaction of PCK1 with PPP2R1A or PPP2CA was analyzed by immunoprecipitation and Western blot analysis as indicated in HepG2 cells (WT), HAT1 KO #1 HepG2 cells, HAT1 KO #1 HepG2 cells reconstituted with wild-type HAT1 or HAT1 mutant (T188A). (G) The interaction of flag-PPP2R1A or flag PPP2R1A mutant (K541R) with PCK1 or PPP2CA was examined by immunoprecipitation and Western blot analysis as indicated in HepG2 cells (WT), HAT1 KO #1 HepG2 cells, HAT1 KO #1 HepG2 cells reconstituted with wild-type HAT1 or HAT1 mutant (T188A). (H–M) HepG2 cells were transfected with plasmids overexpressing HAT1, or co-transfected with plasmids overexpressing HAT1 and PPP2R1A or PPP2R1A mutant (K541E), respectively. (H) The levels of PCK1 pS90, PCK1, HAT1, PPP2R1A, and β -actin were examined by Western blot analysis in the cells. (I) The relative PCK1 activity was detected by the corresponding assay kit in the cells. (J) The nuclear accumulation of SREBP1 was observed by confocal immunofluorescence analysis in the cells. Scale bar, 5 μm. (K) The mRNA expression levels of SREBP1 target genes, including FASN, SCD, ACACA, and GPAM, were analyzed by RT-qPCR in the cells. (L) The relative levels of PEP, citrate, triglyceride, and cholesterol were tested by the corresponding assay kits in the cells cultured in 25 or 0.1 mmol/L glucose conditions for 10 h, respectively. (M) The lipid accumulation was measured by Nile Red staining in the cells cultured in 25 or 0.1 mmol/L glucose conditions for 10 h, respectively. Scale bar, 5 μm. The relative Nile Red staining intensity was quantified. Data are presented as mean ± SD. Student's t -test, ∗∗∗ P < 0.001. HAT1: histone acetyltransferase 1, PCK1: phosphoenolpyruvate carboxykinase 1, pS90: phosphorylation at serine 90, DEN: diethylnitrosamine, CCl 4 : carbon tetrachloride, HCC: hepatocellular carcinoma, WT: wild type, KO: knockout, PPP2R1A: protein phosphatase 2 scaffold subunit alpha, PPP2CA: protein phosphatase 2 catalytic subunit alpha, PEP: phosphoenolpyruvate, SREBP1: sterol regulatory element-binding protein 1, FASN: fatty acid synthase, SCD: stearoyl-CoA desaturase, ACACA: acetyl-CoA carboxylase alpha, GPAM: glycerol-3-phosphate acyltransferase 1.

Article Snippet: Histone acetyltransferase 1 (HAT1) inhibitor JG-2016 was purchased from MCE (Shanghai, China).

Techniques: Phospho-proteomics, Immunoprecipitation, Western Blot, Mutagenesis, In Vitro, Purification, Transfection, Activity Assay, Immunofluorescence, Expressing, Quantitative RT-PCR, Cell Culture, Staining, Knock-Out, Binding Assay

Journal: Acta Pharmaceutica Sinica. B

Article Title: Succinylation of tumor suppressor PPP2R1A K541 by HAT1 converses the role in modulation of gluconeogenesis/lipogenesis remodeling to display oncogene function

doi: 10.1016/j.apsb.2025.07.040

Figure Lengend Snippet: PPP2R1A promotes gluconeogenesis and inhibits lipogenesis by dephosphorylation of PCK1 Ser90 to suppress tumor growth. (A–I) HepG2 cells were transfected with plasmids overexpressing PPP2R1A or PPP2R1A mutant (K541E), or co-transfected with plasmids overexpressing PCK1 or PCK1 mutant (S90A), respectively. (A) The levels of PCK1 pS90, PCK1, PPP2R1A, and β -actin were tested by Western blot analysis in the cells. (B) The relative PCK1 activity was analyzed by the corresponding assay kit in the cells. (C) The nuclear accumulation of SREBP1 was observed by confocal immunofluorescence analysis in the cells. Scale bar, 5 μm. (D) The mRNA expression levels of SREBP1 target genes, including FASN, SCD, ACACA, and GPAM, were examined by RT-qPCR in the cells. (E–H) The relative levels of PEP, citrate, triglyceride, and cholesterol were analyzed by the corresponding assay kits in the cells cultured in 25 or 0.1 mmol/L glucose conditions for 10 h, respectively. (I) The cell viability was determined by CCK-8 assays cultured in 25 or 0.1 mmol/L glucose conditions for 10 h, respectively. (J–N) HepG2 cells were transfected with plasmids overexpressing PPP2R1A or PPP2R1A mutant (K541E), or co-transfected with plasmids overexpressing PCK1 or PCK1 mutant (S90A), respectively. The cells were subcutaneously injected into athymic nude mice ( n = 6). (J) Representative images of tumors from the nude mice ( n = 6) are shown. Tumor volumes and average tumor weight were calculated. Scale bar, 1 cm. (K–N) The relative levels of PEP, citrate, triglyceride, and cholesterol were analyzed by the corresponding assay kits in the tumor tissues from the nude mice. Data are presented as mean ± SD. Student's t -test, ∗∗ P < 0.01, ∗∗∗ P < 0.001. HAT1: histone acetyltransferase 1, PCK1: phosphoenolpyruvate carboxykinase 1, pS90: phosphorylation at serine 90, PPP2R1A: protein phosphatase 2 scaffold subunit alpha, SREBP1: sterol regulatory element-binding protein 1, FASN: fatty acid synthase, SCD: stearoyl-CoA desaturase, ACACA: acetyl-CoA carboxylase alpha, GPAM: glycerol-3-phosphate acyltransferase 1.

Article Snippet: Histone acetyltransferase 1 (HAT1) inhibitor JG-2016 was purchased from MCE (Shanghai, China).

Techniques: De-Phosphorylation Assay, Transfection, Mutagenesis, Western Blot, Activity Assay, Immunofluorescence, Expressing, Quantitative RT-PCR, Cell Culture, CCK-8 Assay, Injection, Phospho-proteomics, Binding Assay

Journal: Acta Pharmaceutica Sinica. B

Article Title: Succinylation of tumor suppressor PPP2R1A K541 by HAT1 converses the role in modulation of gluconeogenesis/lipogenesis remodeling to display oncogene function

doi: 10.1016/j.apsb.2025.07.040

Figure Lengend Snippet: Succinylation of PPP2R1A by HAT1 converses the role in modulation of remodeling of gluconeogenesis/lipogenesis to support liver cancer. (A) The cell viability was analyzed by CCK-8 assays in HepG2 cells (WT), HAT1 KO #1 HepG2 cells, HAT1 KO #1 HepG2 cells reconstituted with wild-type HAT1 or HAT1 mutant (T188A), HAT1 KO #1 HepG2 cells co-reconstituted with wild-type HAT1 and PCK1, PCK1 mutant (S90A), PPP2R1A, or PPP2R1A mutant (K541R), respectively. (B–H) The athymic nude mice ( n = 6) were subcutaneously injected with HepG2 cells (WT), HAT1 KO #1 HepG2 cells, HAT1 KO #1 HepG2 cells reconstituted with wild-type HAT1 or HAT1 mutant (T188A), HAT1 KO #1 HepG2 cells co-reconstituted with wild-type HAT1 and PCK1, PCK1 mutant (S90A), PPP2R1A, or PPP2R1A mutant (K541R), respectively. (B) Representative images of tumors from the nude mice ( n = 6) were shown. Tumor volumes and average tumor weight were calculated. Scale bar = 1 cm. (C) The expression levels of Ki-67 were analyzed by IHC staining in the tumor tissues from the nude mice. Scale bar = 50 μm. (D) The levels of PCK1 pS90, PCK1, PPP2R1A, HAT1, and β -actin were examined by Western blot analysis in the tumor tissues from the nude mice. (E–H) The relative levels of PEP, citrate, triglyceride, and cholesterol were determined by the corresponding assay kits in the tumor tissues from the nude mice. Data are presented as mean ± SD. Student's t -test, ∗∗∗ P < 0.001. HAT1: histone acetyltransferase 1, PCK1: phosphoenolpyruvate carboxykinase 1, pS90: phosphorylation at serine 90, WT: wild type, KO: knockout, PPP2R1A: protein phosphatase 2 scaffold subunit alpha, IHC: immunohistochemistry, PEP: phosphoenolpyruvate.

Article Snippet: Histone acetyltransferase 1 (HAT1) inhibitor JG-2016 was purchased from MCE (Shanghai, China).

Techniques: CCK-8 Assay, Mutagenesis, Injection, Expressing, Immunohistochemistry, Western Blot, Phospho-proteomics, Knock-Out

Journal: Acta Pharmaceutica Sinica. B

Article Title: Succinylation of tumor suppressor PPP2R1A K541 by HAT1 converses the role in modulation of gluconeogenesis/lipogenesis remodeling to display oncogene function

doi: 10.1016/j.apsb.2025.07.040

Figure Lengend Snippet: Succinylation of PPP2R1A lysine 541 by HAT1 converses the role in modulation of remodeling of gluconeogenesis/lipogenesis to support liver cancer. In this model, PP2A is a protein complex that contains 60 different holoenzymes, PPP2R1A and PPP2CA are representative members of A and C subunits of PP2A in hepatocytes. PPP2R1A is required for the assembly of the PP2A complex. PCK1 catalyzes the conversion of OAA to PEP in gluconeogenesis. PPP2R1A promotes gluconeogenesis and inhibits lipogenesis by de-phosphorylation of PCK1 Ser90, leading to the tumor suppression of liver cancer. HAT1 contributes to the remodeling of gluconeogenesis/lipogenesis in liver cancer. PPP2R1A is succinylated at K541 by HAT1, leading to the inhibition of assembly of PP2A in liver cancer. Inhibition of assembly of PP2A blocks the dephosphorylation of PCK1 Ser90, which induces lipogenesis but fails to catalyze the conversion of OAA to PEP in gluconeogenesis, contributing to the malignant progression of liver cancer. Thus, the succinylation of tumor suppressor PPP2R1A displays an oncogene function. HAT1: histone acetyltransferase 1, PCK1: phosphoenolpyruvate carboxykinase 1, PPP2R1A: protein phosphatase 2 scaffold subunit alpha, PPP2CA: protein phosphatase 2 catalytic subunit alpha, PP2A: protein phosphatase 2A, OAA: oxaloacetate, PEP: phosphoenolpyruvate, SREBP1: sterol regulatory element-binding protein 1.

Article Snippet: Histone acetyltransferase 1 (HAT1) inhibitor JG-2016 was purchased from MCE (Shanghai, China).

Techniques: De-Phosphorylation Assay, Inhibition, Binding Assay