Journal: Medical Oncology (Northwood, London, England)

Article Title: Valeric acid acts as a novel HDAC3 inhibitor against prostate cancer

doi: 10.1007/s12032-022-01814-9

Figure Lengend Snippet: VA suppresses the expression of HDAC3. In both PC-3 and DU145 cells, after 48-h treatment of VA, and the expression of HDAC3 was reduced significantly, compared with NC group ( P < 0.05) ( A – E ). Meanwhile, in both PC-3 and DU145 cells, after 48-h treatment of VA, the expression of HDAC3 protein was reduced, compared with NC group ( F )

Article Snippet: For establishing HDAC3-knockout clones, PC-3 and DU145 cells were both conducted using HDAC3 human gene-knockout kit (Crispr) (OriGene Technologies, USA) following the manufacturer’s instruction as described in our previous study [ ].

Techniques: Expressing

Journal: Medical Oncology (Northwood, London, England)

Article Title: Valeric acid acts as a novel HDAC3 inhibitor against prostate cancer

doi: 10.1007/s12032-022-01814-9

Figure Lengend Snippet: Effect of VA on proliferation of prostate cancer cell lines. VA has anti-proliferative effect on prostate cancer cells while being low toxic to normal cells and such effect could be reduced by HDAC3 gene knockout. The inhibition rates of PC‐3 in the presence of 100 μM VA ranged from 34.42 ± 5.9% to 56.08 ± 1.28% during 24 h to 96 h and displayed significantly higher inhibition rates when compared to the 50-μM VA group with the range from 16.52 ± 2.32% to 24.76 ± 2.32% ( P < 0.001, from 24 to 96 h) ( A ). Similar trends have been displayed in DU145 ( B ). Moreover, for RWPE-1 and RWPE-2, the inhibition rates of 50 μM VA group were significantly lower when compared to 100 μM group, respectively ( C and D ). The relative HDAC3 expression was significantly decreased in HDAC3-KO group than none transfection group in both PC-3 and DU145 cell lines ( E ). The inhibition rates were significantly decreased in CAXII-KO group compared to both non-transfection group and none-KO group, at 24, 48, 72, and 96 h, respectively, in PC-3 and DU145 cell lines ( F and G )

Article Snippet: For establishing HDAC3-knockout clones, PC-3 and DU145 cells were both conducted using HDAC3 human gene-knockout kit (Crispr) (OriGene Technologies, USA) following the manufacturer’s instruction as described in our previous study [ ].

Techniques: Gene Knockout, Inhibition, Expressing, Transfection

Journal: Medical Oncology (Northwood, London, England)

Article Title: Valeric acid acts as a novel HDAC3 inhibitor against prostate cancer

doi: 10.1007/s12032-022-01814-9

Figure Lengend Snippet: The representative 3D spheroid models of PC-3 ( A ) and DU145 ( B ) cells were treated by VA and NC, respectively. The cross-section area inhibition rates eventually climbed to 46.77 ± 19.62% and 48.07 ± 19.72% at 96 h for PC-3 and DU145 cells, respectively ( C ). The CASP3 SA (caspase-3 specific activity) has shown that VA evaluated the caspase-3 activity in both PC-3 and DU145 cells cultured in either 2D or 3D system, compared to respective NC ( P < 0.05) ( D ). Relative expression of E2F1 and E2F3 in VA group was significantly lower that of NC, respectively ( P < 0.001) ( E, F ), NC negative control

Article Snippet: For establishing HDAC3-knockout clones, PC-3 and DU145 cells were both conducted using HDAC3 human gene-knockout kit (Crispr) (OriGene Technologies, USA) following the manufacturer’s instruction as described in our previous study [ ].

Techniques: Inhibition, Activity Assay, Cell Culture, Expressing, Negative Control

Journal: Clinical epigenetics

Article Title: Nrf2 status affects tumor growth, HDAC3 gene promoter associations, and the response to sulforaphane in the colon.

doi: 10.1186/s13148-015-0132-y

Figure Lengend Snippet: Fig. 2 SFN decreased HDAC activity and HDAC3 protein expression in mouse colon tumors. a HDAC activity in colon tumor lysates was determined as described in “Methods” section. Mean values were calculated in each group (dotted line). *P < 0.05 vs. no SFN controls. b HDAC expression and histone acetylation determined by immunoblotting. c Densitometry data for HDACs were normalized to β-actin, whereas acetylated histone H4K12 (AcH4K12) was normalized to histone H4. Data = mean ± SD (n = 3); *P < 0.05 vs. no SFN controls

Article Snippet: Cells were transfected for 48 h with HDAC3 siRNA (Trilencer-27, OriGene), Keap1 siRNA (Sigma-Aldrich), control siRNA (OriGene), or Lipofectamine 2000 alone, using the manufacturer’s protocol (Invitrogen).

Techniques: Activity Assay, Expressing, Western Blot

Journal: Clinical epigenetics

Article Title: Nrf2 status affects tumor growth, HDAC3 gene promoter associations, and the response to sulforaphane in the colon.

doi: 10.1186/s13148-015-0132-y

Figure Lengend Snippet: Fig. 7 HDAC3 knockdown induces p16 expression in human colon cancer cells. a HCT116 cells were transfected with non-specific scrambled negative control siRNA (Neg siRNA), HDAC3 siRNA, or Keap1 siRNA for 48 h, followed by vehicle (−SFN) or 15-μM SFN treatment for 24 h. p16 mRNA levels were normalized to ACTB; data = mean ± SD (n = 3), *P < 0.05 compared to untreated Neg siRNA. b The corresponding whole cell lysates were immunoblotted for p16, HDAC3, HDAC6 and Nrf2. HDAC6 was unchanged and served as loading control. c The same samples were analyzed for changes in heme oxygenase-1 (HO1), a known Nrf2 target gene. d–e Confirmation of Keap1 and HDAC3 knockdown. Kelch-like ECH-associated protein 1 (Keap1)

Article Snippet: Cells were transfected for 48 h with HDAC3 siRNA (Trilencer-27, OriGene), Keap1 siRNA (Sigma-Aldrich), control siRNA (OriGene), or Lipofectamine 2000 alone, using the manufacturer’s protocol (Invitrogen).

Techniques: Knockdown, Expressing, Transfection, Negative Control, Control

Journal: Clinical epigenetics

Article Title: Nrf2 status affects tumor growth, HDAC3 gene promoter associations, and the response to sulforaphane in the colon.

doi: 10.1186/s13148-015-0132-y

Figure Lengend Snippet: Fig. 6 Nrf2 status affects p16 expression and HDAC3 promoter interactions in mouse colon tumors. a p16 mRNA expression levels in mouse colon tumors were normalized to Actb. Data = mean ± SD (n = 3); *P < 0.05 for control diet vs. SFN treatment. b–e Representative immunohistochemical staining (×10) of p16 in mouse colon tumor and adjacent normal colon; tumor (T), normal (N). A magnified (×40) image is shown for each dotted box, with a 50-μm scale bar included in b, inset. f In vivo ChIP assays on mouse colon tumors, interrogating HDAC3 interactions on p16. Data = mean ± SD and are representative of three independent experiments. A region 10 Kb upstream of the p16 promoter, containing a Runx1 binding site, served as a negative control for the ChIP assays

Article Snippet: Cells were transfected for 48 h with HDAC3 siRNA (Trilencer-27, OriGene), Keap1 siRNA (Sigma-Aldrich), control siRNA (OriGene), or Lipofectamine 2000 alone, using the manufacturer’s protocol (Invitrogen).

Techniques: Expressing, Control, Immunohistochemical staining, Staining, In Vivo, Binding Assay, Negative Control

Journal: Clinical epigenetics

Article Title: Nrf2 status affects tumor growth, HDAC3 gene promoter associations, and the response to sulforaphane in the colon.

doi: 10.1186/s13148-015-0132-y

Figure Lengend Snippet: Fig. 8 Human subjects given a broccoli sprout extract, or consuming high levels of cruciferous vegetables, have altered HDAC3 and p16 expression. a Human subjects (n = 5, each arm of the study) consumed a broccoli sprout extract supplement (BSE, 200 μmol SFN equivalent, IND #111736) or a placebo for 7 days, and blood was drawn at the times indicated. b Total levels of SFN and its metabolites in plasma (black symbols), and b, c HDAC3 and p16 protein expression changes in circulating PBMCs. Results are from a single volunteer and are representative of other subjects who took the BSE supplement. No such changes occurred after taking the placebo (data not shown). Individual SFN metabolites, noted in Additional file 3: Table S2, were determined as reported [27]. d Study design in screening colonoscopy patients. e Relative mRNA expression of p16 in subjects reporting low (0–1 servings/week) vs. high (>5 servings/week) cruciferous vegetable intake. f Immunoblotting of colon biopsies, arranged left to right according to cruciferous vegetable intake. g–i Associations between p16, HDAC3, AcH4K12, and Nrf2 normalized to β-actin (relative densitometry of proteins in panel f)

Article Snippet: Cells were transfected for 48 h with HDAC3 siRNA (Trilencer-27, OriGene), Keap1 siRNA (Sigma-Aldrich), control siRNA (OriGene), or Lipofectamine 2000 alone, using the manufacturer’s protocol (Invitrogen).

Techniques: Expressing, Clinical Proteomics, Western Blot