human hdac4  (Vector Biolabs)

Journal: Arteriosclerosis, thrombosis, and vascular biology

Article Title: Focal adhesion kinase promotes calcification of vascular smooth muscle cells via regulation of histone deacetylase 4 and 5

doi: 10.1161/ATVBAHA.123.319010

Figure Lengend Snippet: A, VS-6063 reduced levels of phosphorylated HDAC4 and HDAC5. n=4 biological replicates in each group. B, Silencing FAK also reduced the phosphorylation of HDAC4 and HDAC5 in HASMCs grown in common medium or osteogenic medium. n=4 biological replicates in each group. C, FAK inhibition using VS6063 decreased cytosolic localization and increased nuclear localization of HDAC4 and HDAC5 in osteogenic media as shown by immunofluorescence images (60x). 30 cells in each group were used for analysis.

Article Snippet: Recombinant adenoviruses expressing human HDAC4 or HDAC5 were obtained from Vector Biolabs (HDAC4: #1435; HDAC5: #210890).

Techniques: Inhibition, Phospho-proteomics, Immunofluorescence

Journal: Arteriosclerosis, thrombosis, and vascular biology

Article Title: Focal adhesion kinase promotes calcification of vascular smooth muscle cells via regulation of histone deacetylase 4 and 5

doi: 10.1161/ATVBAHA.123.319010

Figure Lengend Snippet: Leptomycin B (LEP) inhibited nuclear export of HDAC4 and HDAC5 in A , normal media and B , in osteogenic media as shown by immunofluorescence. 12–46 cells in each group were used for analysis. C, Leptomycin B reduced phosphorylated HDAC4 and HDAC5 and inhibited the expression of RUNX2 and ALPL in osteogenic media. n=4 biological replicates in each group.

Article Snippet: Recombinant adenoviruses expressing human HDAC4 or HDAC5 were obtained from Vector Biolabs (HDAC4: #1435; HDAC5: #210890).

Techniques: Inhibition, Immunofluorescence, Expressing

Journal: Arteriosclerosis, thrombosis, and vascular biology

Article Title: Focal adhesion kinase promotes calcification of vascular smooth muscle cells via regulation of histone deacetylase 4 and 5

doi: 10.1161/ATVBAHA.123.319010

Figure Lengend Snippet: A , Increased expression of HDAC4/5 was achieved by adenovirus transduction (Ad.HDAC4 or Ad.HDAC5). B, Ad.HDAC4 or Ad.HDAC5 resulted in enhanced calcification that was inhibited with the treatment of leptomycin B (5nM) n=3 biological replicates in each group (with 2 representative replicates shown). Quantitative calcium assay in cells treated with C, Ad.HDAC4 and D , Ad.HDAC5 in the presence or absence of leptomycin demonstrated reduced calcification with leptomycin treatment. n=3 biological replicates in each group. E and F, Cytosolic and nuclear localization of HDAC4 and HDAC5 in AdHDAC4 and AdHDAC5 treated cells in the presence and absence of leptomycin (a nuclear export inhibitor, 10 nM for 3 hours) in osteogenic media. 12–13 cells in each group were used for analysis.

Article Snippet: Recombinant adenoviruses expressing human HDAC4 or HDAC5 were obtained from Vector Biolabs (HDAC4: #1435; HDAC5: #210890).

Techniques: Over Expression, Expressing, Transduction, Calcium Assay

Journal: Arteriosclerosis, thrombosis, and vascular biology

Article Title: Focal adhesion kinase promotes calcification of vascular smooth muscle cells via regulation of histone deacetylase 4 and 5

doi: 10.1161/ATVBAHA.123.319010

Figure Lengend Snippet: A, Treatment of HASMCs with siHDAC4 resulted in >70% and >65% knockdown of HDAC4 mRNA level in common or osteogenic medium, respectively. n=6 biological replicates in each group. B, Protein levels of RUNX2 and ALPL were increased with osteogenic medium. However, siHDAC4 decreased the levels of RUNX2 and ALPL induced by osteogenic medium. n=4 biological replicates in each group. C, Treatment of HASMCs with siHDAC5 resulted in >55% and >50% knockdown of HDAC5 mRNA level in normal or osteogenic medium, respectively, and decreased HDAC5 protein levels in siHDAC5-treated cells. n=6 biological replicates in each mRNA group. D, siHDAC5 decreased the protein levels of RUNX2 and ALPL induced by osteogenic medium. n=4 biological replicates in each group. E, Treatment with siHDAC4, siHDAC5, or the combination inhibited calcification of HASMCs grown in osteogenic medium for 14 days, as evidenced by Alizarin Red staining. n=3 biological replicates in each group (with 2 representative replicates shown). F, Treatment of HASMCs with LMK-235 (a pharmacologic inhibitor of HDAC4 and HDAC5) inhibited calcification induced by osteogenic medium in a dose-dependent manner. n=2 biological replicates in each group G, LMK-235 reduced the migration of VSMCs induced by osteogenic medium. The experiments in figure 1H and 7G were performed at the same time. n=6 biological replicates in each group.

Article Snippet: Recombinant adenoviruses expressing human HDAC4 or HDAC5 were obtained from Vector Biolabs (HDAC4: #1435; HDAC5: #210890).

Techniques: Inhibition, Knockdown, Staining, Migration

Journal: Arteriosclerosis, thrombosis, and vascular biology

Article Title: Focal adhesion kinase promotes calcification of vascular smooth muscle cells via regulation of histone deacetylase 4 and 5

doi: 10.1161/ATVBAHA.123.319010

Figure Lengend Snippet: A and B, Increased HDAC4 or HDAC5 expression by adenovirus resulted in augmented calcification of HASMCs that was inhibited by treatment with the FAK inhibitor VS-6063 (1μM). n=3 biological replicates in each group (with 2 representative replicates shown). C and D, Reduction of FAK expression with siFAK significantly attenuated the calcification of HASMCs induced by Ad.HDAC4 or Ad.HDAC5 in osteogenic medium or induced by osteogenic medium alone. n=3 biological replicates in each group (with 2 representative replicates shown). E and F, Osteogenic medium induced calcification of mouse aortas and human carotid arteries after culturing for 21 days. However, the calcification induced by osteogenic medium was inhibited by treatment with VS-6063 (2μM or 4μM). n=4 biological replicates in each group.

Article Snippet: Recombinant adenoviruses expressing human HDAC4 or HDAC5 were obtained from Vector Biolabs (HDAC4: #1435; HDAC5: #210890).

Techniques: Inhibition, Over Expression, Cell Culture, Expressing

Journal: Arteriosclerosis, thrombosis, and vascular biology

Article Title: Focal adhesion kinase promotes calcification of vascular smooth muscle cells via regulation of histone deacetylase 4 and 5

doi: 10.1161/ATVBAHA.123.319010

Figure Lengend Snippet: Localization of PTK2 , HDAC5 , and HDAC4 gene expression in modulated SMC subtypes using an integrated human atherosclerosis reference. Uniform Manifold Approximation and Projection (UMAP) embeddings from an integrated human atherosclerosis single-cell RNA-seq reference dataset (see Methods ), highlighting ( A ) PTK2 , ( B ) HDAC5 , and ( C ) HDAC4 normalized gene expression. Individual sequencing libraries across four studies were harmonized after QC and batch correction with reciprocal PCA (rPCA). A broad SMC cluster was annotated using transfer learning with cell labels from the Tabula Sapiens vasculature subset. SMC subtypes were further annotated by extracting gene modules from a scRNA meta-analysis of murine SMCs (contractile SMC, transitional SMC, fibromyocyte, and fibrochondrocyte) and calculating their enrichment in cells within the main SMC cluster. PTK2 and HDAC5 expression were enriched in transitional SMCs and fibromyocytes.

Article Snippet: Recombinant adenoviruses expressing human HDAC4 or HDAC5 were obtained from Vector Biolabs (HDAC4: #1435; HDAC5: #210890).

Techniques: Gene Expression, RNA Sequencing, Sequencing, Expressing

Journal: Journal of Medicinal Chemistry

Article Title: Discovery of Pyrazole-Based Positron Emission Tomography Agent that Maps Histone Deacetylase 6 (HDAC6) in the Nonhuman Primate Brain

doi: 10.1021/acs.jmedchem.5c02216

Figure Lengend Snippet: Representative HDAC6 PET tracers with IC 50 values for HDAC6.

Article Snippet: Recombinant human HDAC6, HDAC7, and HDAC1 were obtained from SignalChem Pharmaceutical Inc. (Richmond, BC, Canada), recombinant HDAC8 was obtained from Reaction Biology Corporation (Marvern, PA, USA), and recombinant human HDAC4 and mouse HDAC6 were prepared by Axcelead Drug Discovery Partners Inc. A test compound (1 μL) diluted with DMSO was added to 75 μL of assay buffer (24 mM Tris-HCl [pH 7.5], 135 mM NaCl, 0.35 mM KCl, 1 mM MgCl 2 , 0.01% Tween 20, 0.6 mM glutathione) in 384 well plates.

Techniques:

Journal: Journal of Medicinal Chemistry

Article Title: Discovery of Pyrazole-Based Positron Emission Tomography Agent that Maps Histone Deacetylase 6 (HDAC6) in the Nonhuman Primate Brain

doi: 10.1021/acs.jmedchem.5c02216

Figure Lengend Snippet: Affinity selection (AS)–MS binding study of HDAC6 with 16a and bavarostat. Data points are the mean ± standard error (SE) (four technical replicates). (A) Saturation binding curve. (B) Time course for the dissociation. (C) Binding parameters calculated from AS–MS data. Functional IC 50 values are shown for comparison.

Article Snippet: Recombinant human HDAC6, HDAC7, and HDAC1 were obtained from SignalChem Pharmaceutical Inc. (Richmond, BC, Canada), recombinant HDAC8 was obtained from Reaction Biology Corporation (Marvern, PA, USA), and recombinant human HDAC4 and mouse HDAC6 were prepared by Axcelead Drug Discovery Partners Inc. A test compound (1 μL) diluted with DMSO was added to 75 μL of assay buffer (24 mM Tris-HCl [pH 7.5], 135 mM NaCl, 0.35 mM KCl, 1 mM MgCl 2 , 0.01% Tween 20, 0.6 mM glutathione) in 384 well plates.

Techniques: Selection, Binding Assay, Functional Assay, Comparison

Journal: Journal of Medicinal Chemistry

Article Title: Discovery of Pyrazole-Based Positron Emission Tomography Agent that Maps Histone Deacetylase 6 (HDAC6) in the Nonhuman Primate Brain

doi: 10.1021/acs.jmedchem.5c02216

Figure Lengend Snippet: Brain and plasma concentration of 16a in wild-type (WT) and HDAC6 knockout (KO) mice. Compound 16a (1 mg/kg) was intravenously administered to WT and HDAC6 KO mice. Two hours after administration, the hippocampus and plasma were collected and the concentration of 16a in the brain (A) and plasma (B) was measured using liquid chromatography with tandem mass spectrometry (LC–MS/MS). The data are expressed in percentage of injected dose per gram of brain tissue (%ID/g) or per milliliter of plasma (%ID/mL), respectively, as mean ± standard error (SE), n = 4. ** P < 0.01, n.s.: not significant.

Article Snippet: Recombinant human HDAC6, HDAC7, and HDAC1 were obtained from SignalChem Pharmaceutical Inc. (Richmond, BC, Canada), recombinant HDAC8 was obtained from Reaction Biology Corporation (Marvern, PA, USA), and recombinant human HDAC4 and mouse HDAC6 were prepared by Axcelead Drug Discovery Partners Inc. A test compound (1 μL) diluted with DMSO was added to 75 μL of assay buffer (24 mM Tris-HCl [pH 7.5], 135 mM NaCl, 0.35 mM KCl, 1 mM MgCl 2 , 0.01% Tween 20, 0.6 mM glutathione) in 384 well plates.

Techniques: Clinical Proteomics, Concentration Assay, Knock-Out, Liquid Chromatography, Mass Spectrometry, Liquid Chromatography with Mass Spectroscopy, Injection

Journal: Journal of Medicinal Chemistry

Article Title: Discovery of Pyrazole-Based Positron Emission Tomography Agent that Maps Histone Deacetylase 6 (HDAC6) in the Nonhuman Primate Brain

doi: 10.1021/acs.jmedchem.5c02216

Figure Lengend Snippet: Comparison of HDAC6 expression levels in a mouse and monkey using Western blotting. Hippocampus (Hip) was obtained from two mice, and HDAC6 expression was examined. Recombinant HDAC6 conjugated with GST (0.03, 0.1, 0.3, and 1 ng) was also used for the positive controls (A). Lysates of the hippocampus and corpus callosum (CC) obtained from a monkey were applied to Western blotting in duplicate. The same controls were used for comparison (B).

Article Snippet: Recombinant human HDAC6, HDAC7, and HDAC1 were obtained from SignalChem Pharmaceutical Inc. (Richmond, BC, Canada), recombinant HDAC8 was obtained from Reaction Biology Corporation (Marvern, PA, USA), and recombinant human HDAC4 and mouse HDAC6 were prepared by Axcelead Drug Discovery Partners Inc. A test compound (1 μL) diluted with DMSO was added to 75 μL of assay buffer (24 mM Tris-HCl [pH 7.5], 135 mM NaCl, 0.35 mM KCl, 1 mM MgCl 2 , 0.01% Tween 20, 0.6 mM glutathione) in 384 well plates.

Techniques: Comparison, Expressing, Western Blot, Recombinant

Journal: Journal of Medicinal Chemistry

Article Title: Discovery of Pyrazole-Based Positron Emission Tomography Agent that Maps Histone Deacetylase 6 (HDAC6) in the Nonhuman Primate Brain

doi: 10.1021/acs.jmedchem.5c02216

Figure Lengend Snippet: Immunohistochemical (IHC) staining of HDAC6 protein in a monkey brain. Coronal brain sections, including the areas below were prepared from a monkey (a, putamen; b, caudate nucleus; c, corpus callosum; d, white matter; e, anterior cingulate cortex; f, thalamus; g, hippocampus; h, cerebellum; i, brain stem [pons]). They were immunostained with HDAC6 antibody.

Article Snippet: Recombinant human HDAC6, HDAC7, and HDAC1 were obtained from SignalChem Pharmaceutical Inc. (Richmond, BC, Canada), recombinant HDAC8 was obtained from Reaction Biology Corporation (Marvern, PA, USA), and recombinant human HDAC4 and mouse HDAC6 were prepared by Axcelead Drug Discovery Partners Inc. A test compound (1 μL) diluted with DMSO was added to 75 μL of assay buffer (24 mM Tris-HCl [pH 7.5], 135 mM NaCl, 0.35 mM KCl, 1 mM MgCl 2 , 0.01% Tween 20, 0.6 mM glutathione) in 384 well plates.

Techniques: Immunohistochemical staining, Immunohistochemistry

Journal: British journal of pharmacology

Article Title: A novel inhibitor of class IIa histone deacetylases attenuates collagen-induced arthritis.

doi: 10.1111/bph.17306

Figure Lengend Snippet: FIGURE 2 IL6 up-regulates protein expression and activity of histone deacetylases (HDACs) 4, 5 and 7 but not HDAC1 in HMDMs. (a– d) Representative immunoblot analysis measuring levels of the class I HDAC HDAC1 (a), class IIa HDACs HDAC4 (b), HDAC5 (c) and HDAC7 (d) in IL6-stimulated HMDMs at 4 h or 24 h. Data were normalised to matched control and GAPDH was used as loading control. (e) Cellular class IIa HDAC enzymatic activity in HMDMs as measured using a class IIa HDAC-selective substrate. HMDMs were pre-treated with LL87 (5 μM) for 1 h followed by IL6 (10 ngml1) stimulation for 24 h. Class IIa HDAC activity was measured as the change in relative fluorescence units (ΔRFU = RFUsample RFUblank). Data are presented as mean ± SEM; n = 5 donors. Analysis by nonparametric Kruskal-Wallis test followed by Dunn's multiple comparisons test, * P < 0.05; ns, no significance.

Article Snippet: For class II HDACs (HDAC4, 5, 7, 9, 6), various concentrations of test compounds were pre-incubated with human HDAC4 (5 ng ml 1, BPS Bioscience), HDAC5 (5 ng ml 1, Reaction Biology), HDAC7 F IGURE 1 A novel inhibitor of class IIa HDACs with anti-inflammatory activity in macrophages. (a) Structure of LL87. (b) Concentrationdependent cytotoxicity in HMDMs (MTT cell viability) after 72 h incubation with histone deacetylase (HDAC) inhibitors at various concentrations (M) versus the JAK inhibitor baricitinib. (c) Comparison of LL87 versus baricitinib for inhibition of LPS-induced secretion of cytokines from HMDMs.

Techniques: Expressing, Activity Assay, Western Blot, Control, Fluorescence

Journal: International Journal of Biological Sciences

Article Title: Deacetylation of Glutaminase by HDAC4 contributes to Lung Cancer Tumorigenesis

doi: 10.7150/ijbs.69882

Figure Lengend Snippet: HDAC4 is responsible for deacetylation of GAC at K311. A, B Indicated plasmids were transfected into H1299 cells. Interaction between HDAC4 and GAC was detected by immunoprecipitation and western blot. WCL: whole cell lysate. C Indicated plasmids were transfected into H1299 cells. The protein expression was determined by western blot and glutaminase activity assay was performed. D Indicated siRNAs were transfected into H1299 cells. The protein expression was determined by western blot and glutaminase activity assay was performed. E Indicated plasmids and siRNAs were transfected into H1299 cells and glutaminase activity assay was performed. F The mitochondrial and cytosolic proteins in H1299 cells were separated and the location of HDAC4 and GAC was determined by western blot. VDAC was used as a marker of mitochondrial proteins and GAPDH was used as a marker of cytosolic proteins. Data are showed as mean ± SD, n=3. ** P < 0.01, ns P >0.05.

Article Snippet: The HDAC4 siRNAs (OriGene, SR306523) and TRIM21 siRNAs (OriGene, SR304594) were purchased from OriGene.

Techniques: Transfection, Immunoprecipitation, Western Blot, Expressing, Activity Assay, Marker

Journal: International Journal of Biological Sciences

Article Title: Deacetylation of Glutaminase by HDAC4 contributes to Lung Cancer Tumorigenesis

doi: 10.7150/ijbs.69882

Figure Lengend Snippet: HDAC4 promoted cell proliferation and migration in NSCLC cells. A, B Indicated plasmids were transfected into H1299 (A) and A549 (B) cells and cell growth assay was performed. C, D Indicated siRNAs were transfected into H1299 (C) and A549 (D) cells and cell growth assay was performed. E, F Indicated siRNAs and plasmids were transfected into H1299 (E) and A549 (F) cells and cell growth assay was performed. Western blot assay was performed to confirm the transfection efficiency. G, H Indicated siRNAs were transfected into H1299 cells (G) and H292 cells (H) and cell wound healing assay was performed (scale bar: 500 µm, magnification: 100×). I F-actin staining assay. Indicated siRNAs were transfected into H1299 cells. 48 h later, cells were stained with phalloidin and DAPI. Scale bar=20 µm. Data are showed as mean ± SD, n=3. **P<0.01, ***P<0.001.

Article Snippet: The HDAC4 siRNAs (OriGene, SR306523) and TRIM21 siRNAs (OriGene, SR304594) were purchased from OriGene.

Techniques: Migration, Transfection, Growth Assay, Western Blot, Wound Healing Assay, Staining

Journal: International Journal of Biological Sciences

Article Title: Deacetylation of Glutaminase by HDAC4 contributes to Lung Cancer Tumorigenesis

doi: 10.7150/ijbs.69882

Figure Lengend Snippet: A working model of GAC acetylation in NSCLC. A working model depicting the molecular mechanism of HDAC4 mediated GAC deacetylation and TRIM21 mediated GAC ubiquitination to regulation tumorigenesis in NSCLC.

Article Snippet: The HDAC4 siRNAs (OriGene, SR306523) and TRIM21 siRNAs (OriGene, SR304594) were purchased from OriGene.

Techniques:

Journal: International Journal of Biological Sciences

Article Title: Deacetylation of Glutaminase by HDAC4 contributes to Lung Cancer Tumorigenesis

doi: 10.7150/ijbs.69882

Figure Lengend Snippet: TRIM21 is the E3 ligase for GAC. A, B Indicated plasmids were transfected into H1299 cells. Interaction between GAC and TRIM21 were detected by immunoprecipitation and western blot assay. C-E Indicated plasmids were transfected into H1299 cells. The levels of GAC ubiquitination were detected by immunoprecipitation and western blot assay. F-H Indicated plasmids and siRNAs were transfected into H1299 cells. The levels of GAC ubiquitination were detected by immunoprecipitation and western blot assay. WCL: whole cell lysate. I Indicated siRNAs were transfected into H1299 cells. The protein expression was determined by western blot and glutaminase activity assay was performed. J Indicated plasmids were transfected into H1299 cells. The protein expression were determined by western blot and glutaminase activity assay was performed. Data are showed as mean ± SD, n=3. ** P < 0.01.

Article Snippet: The HDAC4 siRNAs (OriGene, SR306523) and TRIM21 siRNAs (OriGene, SR304594) were purchased from OriGene.

Techniques: Transfection, Immunoprecipitation, Western Blot, Expressing, Activity Assay

Journal: International Journal of Biological Sciences

Article Title: Deacetylation of Glutaminase by HDAC4 contributes to Lung Cancer Tumorigenesis

doi: 10.7150/ijbs.69882

Figure Lengend Snippet: Lys311 acetylation promotes GAC-TRIM21 interaction and GAC ubiquitination. A Indicated plasmids were transfected into H1299 cells. Interaction between GAC and TRIM21 were detected by immunoprecipitation and western blot assay. B Indicated plasmids were transfected into H1299 cells followed by treatment with or without NAM and TSA. Interaction between GAC and TRIM21 were detected by immunoprecipitation and western blot assay. C Indicated plasmids were transfected into H1299 cells. Interaction between GAC and TRIM21 were detected by immunoprecipitation and western blot assay. D Indicated plasmids were transfected into H1299 cells. Interaction between GAC and TRIM21 were detected by immunoprecipitation and western blot assay. E Indicated plasmids were transfected into H1299 cells. The levels of GAC ubiquitination were detected by immunoprecipitation and western blot assay. WCL: whole cell lysate.

Article Snippet: The HDAC4 siRNAs (OriGene, SR306523) and TRIM21 siRNAs (OriGene, SR304594) were purchased from OriGene.

Techniques: Transfection, Immunoprecipitation, Western Blot

Journal: International Journal of Biological Sciences

Article Title: Deacetylation of Glutaminase by HDAC4 contributes to Lung Cancer Tumorigenesis

doi: 10.7150/ijbs.69882

Figure Lengend Snippet: A working model of GAC acetylation in NSCLC. A working model depicting the molecular mechanism of HDAC4 mediated GAC deacetylation and TRIM21 mediated GAC ubiquitination to regulation tumorigenesis in NSCLC.

Article Snippet: The HDAC4 siRNAs (OriGene, SR306523) and TRIM21 siRNAs (OriGene, SR304594) were purchased from OriGene.

Techniques:

Journal: Cellular and molecular biology (Noisy-le-Grand, France)

Article Title: Regulations of LINC0196/miR-584-5p/miR-34a-5p/TRIM59 on Progression of Pediatric Neuroblastoma.

doi: 10.14715/cmb/2022.68.6.19

Figure Lengend Snippet: Figure 2. Comparison of the expressions of LINC0196, miR-584-5p, miR-34a-5p, and TRIM59 in SK-N-SH cells in each group.

Article Snippet: Rabbit anti-human TRIM59 antibody and rabbit anti-human phosphorylated TRIM59 were purchased from Cell Signaling Technology, Inc., the United States. small interfering- (si-) LINC0196, pcDNA-LINC0196, miR-584-5p/miR-34a-5p, and miR-584-5p/miR-34a-5p inhibitors were from Beijing Zhong Shan -Golden Bridge Biological Technology CO., LTD. CCK-8 cell proliferation detection kits and related detection kits of real-time quantitative polymerase chain reaction (RT-qPCR) were of LightCycler 480 products, Roche Molecular Systems, Inc., Switzerland.

Techniques: Comparison

Journal: Cellular and molecular biology (Noisy-le-Grand, France)

Article Title: Regulations of LINC0196/miR-584-5p/miR-34a-5p/TRIM59 on Progression of Pediatric Neuroblastoma.

doi: 10.14715/cmb/2022.68.6.19

Figure Lengend Snippet: Figure 1. Expression of LINC0196, miR-584-5p, miR-34a-5p, and TRIM59 in normal tissues and SK-N-SH cells.

Article Snippet: Rabbit anti-human TRIM59 antibody and rabbit anti-human phosphorylated TRIM59 were purchased from Cell Signaling Technology, Inc., the United States. small interfering- (si-) LINC0196, pcDNA-LINC0196, miR-584-5p/miR-34a-5p, and miR-584-5p/miR-34a-5p inhibitors were from Beijing Zhong Shan -Golden Bridge Biological Technology CO., LTD. CCK-8 cell proliferation detection kits and related detection kits of real-time quantitative polymerase chain reaction (RT-qPCR) were of LightCycler 480 products, Roche Molecular Systems, Inc., Switzerland.

Techniques: Expressing