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Link to original content: http://pubmed.ncbi.nlm.nih.gov/38805168/
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. 2024 Jul;46(7):871-879.
doi: 10.1007/s13258-024-01524-8. Epub 2024 May 28.

Negative regulation of HDAC3 transcription by histone acetyltransferase TIP60 in colon cancer

Seong Yun Lee #  1 , Junyoung Park #  1 , Sang Beom Seo  2
Affiliations

Negative regulation of HDAC3 transcription by histone acetyltransferase TIP60 in colon cancer

Seong Yun Lee et al. Genes Genomics. 2024 Jul.

Abstract

Background: Colon cancer is the third most common cancer globally. The expression of histone deacetylase 3 (HDAC3) is upregulated, whereas the expression of tat interactive protein, 60 kDa (TIP60) is downregulated in colon cancer. However, the relationship between HDAC3 and TIP60 in colon cancer has not been clearly elucidated.

Objective: We investigated whether TIP60 could regulate the expression of HDAC3 and suppress colon cancer cell proliferation.

Methods: RNA sequencing data (GSE108834) showed that HDAC3 expression was regulated by TIP60. Subsequently, we generated TIP60-knockdown HCT116 cells and examined the expression of HDAC3 by western blotting and reverse transcription-quantitative polymerase chain reaction (RT-qPCR). We examined the expression pattern of HDAC3 in various cancers using publicly available datasets. The promoter activity of HDAC3 was validated using a dual-luciferase assay, and transcription factors binding to HDAC3 were identified using GeneCards and Promo databases, followed by validation using chromatin immunoprecipitation-quantitative polymerase chain reaction. Cell proliferation and apoptosis were assessed using colony formation assays and fluorescence-activated cell sorting analysis of HCT116 cell lines.

Results: In response to TIP60 knockdown, the expression level and promoter activity of HDAC3 increased. Conversely, when HDAC3 was downregulated by overexpression of TIP60, proliferation of HCT116 cells was inhibited and apoptosis was promoted.

Conclusion: TIP60 plays a crucial role in the regulation of HDAC3 transcription, thereby influencing cell proliferation and apoptosis in colon cancer. Consequently, TIP60 may function as a tumor suppressor by inhibiting HDAC3 expression in colon cancer cells.

Keywords: HDAC3; Colon cancer; TIP60; Transcription.

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Conflict of interest statement

The authors declare no conflicts of interest.

Figures

Fig. 1
Fig. 1
TIP60 regulated the expression of HDACs in HCT116 cells. A Volcano plot representing RNA sequencing (RNA-seq) analysis results for GSE108834. Red lines indicate the thresholds for differentially expressed genes (DEGs) (|log2 fold change|> 1 and P < 0.05). DEG analysis was performed using DESeq2. B Expression of histone deacetylases (HDACs) in HCT116 cells transfected with siTIP60 (siTIP60 HCT116 cells) according to the RNA-seq results shown in panel (A). *P < 0.05, **P < 0.01 and ****P < 0.0001. C The bar graph represents reverse transcription-quantitative polymerase chain reaction (RT-qPCR) analysis to determine the mRNA levels of HDAC1 and HDAC2 in TIP60-knockdown (shTIP60) HCT116 cells. Data are shown as the mean ± standard error of the mean (SEM; n = 3). P-values were calculated using paired two-tailed Student’s t-tests. D The bar graph represents the RT-qPCR analysis of the mRNA levels of HDAC3 and HDAC4 in shTIP60 HCT116 cells. Data are shown as the mean ± SEM (n = 3). P-values were calculated using paired two-tailed Student’s t-tests. *P < 0.05 and ***P < 0.001. E Western blot analysis to evaluate HDAC1 and HDAC2 protein level in shTIP60 HCT116 cells. F Western blot analysis to evaluate HDAC3 and HDAC4 protein level in shTIP60 HCT116 cells
Fig. 2
Fig. 2
HDAC3 was upregulated in various cancers. A Box plot showing the expression level of HDAC3 in colon adenocarcinoma (COAD) and rectal adenocarcinoma (READ) compared with corresponding normal tissue. Data were analyzed using the GEPIA2 database. B Box plot showing the expression level of HDAC3 in breast invasive carcinoma (BRCA) compared to the corresponding normal tissue. Data were analyzed using the GEPIA2 database. C Box plot showing the expression level of HDAC3 in uterine corpus endometrial carcinoma (UCEC) compared with the corresponding normal tissue. Data were analyzed using the GEPIA2 database. D Gene ontology (GO) analysis result of positively correlated genes with HDAC3 in colon cancer. The positively correlated genes were identified using UALCAN and GO analysis was performed using ShinyGO. E GO analysis result of positively correlated genes with HDAC3 in rectum cancer. The positively correlated genes were identified using UALCAN and GO analysis was performed using ShinyGO
Fig. 3
Fig. 3
TIP60 acted as a transcriptional repressor of HDAC3 in HCT116 cells. A Dual-luciferase assay to evaluate the promoter activity of HDAC3 after TIP60 depletion in HCT116 cells. Data are shown as the mean ± SEM (n = 3). P-values were calculated using paired two-tailed Student’s t-tests. *P < 0.05. B Dual-luciferase assay to evaluate promoter activity of HDAC3 after TIP60 overexpression in HCT116 cells. Data are shown as the mean ± SEM (n = 3). P-values were calculated using one-way analysis of variance (ANOVA) followed by Dunnett’s multiple comparison test. *P < 0.05 and **P < 0.01. C The bar graph represents the RT-qPCR analysis of the mRNA levels of HDAC3 target genes in shTIP60 HCT116 cells. Data are shown as the mean ± SEM (n = 3). P-values were calculated using paired two-tailed Student’s t-tests. *P < 0.05, ***P < 0.001, and ****P < 0.0001. D The bar graph represents the RT-qPCR analysis of the mRNA levels of HDAC3 target genes in TIP60-overexpressing HCT116 cells. Data are shown as the mean ± SEM (n = 3). P-values were calculated using paired two-tailed Student’s t-tests. *P < 0.05. E Transcription factor analysis was performed to identify the transcription factors that bind to the HDAC3 promoter region using Promo and GeneCards databases. F Chromatin immunoprecipitation-qPCR (ChIP-qPCR) assay to evaluate occupancies of TIP60 and JUNB at the HDAC3 promoter region in TIP60 knockdown cells. A specific primer set targeting the HDAC3 promoter region was used. Data are shown as the mean ± SEM (n = 3). P-values were calculated using paired two-tailed Student’s t-tests. *P < 0.05 and ***P < 0.001
Fig. 4
Fig. 4
HDAC3 suppression by TIP60 regulated the proliferation and apoptosis of HCT116 cells. A (Left) Representative images of colony formation assay of HCT116 cells in each condition. (Right) The bar graph represents the result of colony formation assay. Data are shown as the mean ± SEM (n = 3). P-values were calculated using ANOVA followed by Dunnett’s multiple comparison test. *P < 0.05 and n.s., not significant. B The bar graph represents RT-qPCR analysis of the mRNA levels of apoptosis-related genes in TIP60 overexpressing HCT116 cells. Data are shown as the mean ± SEM (n = 3). P-values were calculated using paired two-tailed Student’s t-tests. *P < 0.05. C The proportion of apoptotic cells were analyzed by flow cytometry. Transfected cells were treated with 5 mM of hydroxyurea (HU) for 32 h to induce apoptosis and stained with propidium iodide (PI) and FITC-Annexin V after trypsinization. D Western blot analysis to evaluate apoptosis-related protein levels including PARP1 and Bcl-XL in each condition. Asterisk marks (*) indicate cleaved form of PARP1
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References

    1. Adhikari N, Amin SA, Trivedi P, Jha T, Ghosh B. HDAC3 is a potential validated target for cancer: An overview on the benzamide-based selective HDAC3 inhibitors through comparative SAR/QSAR/QAAR approaches. Eur J Med Chem. 2018;157:1127–1142. doi: 10.1016/j.ejmech.2018.08.081. - DOI - PubMed
    1. Avvakumov N, Cote J. The MYST family of histone acetyltransferases and their intimate links to cancer. Oncogene. 2007;26:5395–5407. doi: 10.1038/sj.onc.1210608. - DOI - PubMed
    1. Berger SL. The complex language of chromatin regulation during transcription. Nature. 2007;447:407–412. doi: 10.1038/nature05915. - DOI - PubMed
    1. Castro-Mondragon JA, Riudavets-Puig R, Rauluseviciute I, Lemma RB, Turchi L, Blanc-Mathieu R, Lucas J, Boddie P, Khan A, Manosalva Perez N, et al. JASPAR 2022: the 9th release of the open-access database of transcription factor binding profiles. Nucleic Acids Res. 2022;50:D165–D173. doi: 10.1093/nar/gkab1113. - DOI - PMC - PubMed
    1. Chandrashekar DS, Karthikeyan SK, Korla PK, Patel H, Shovon AR, Athar M, Netto GJ, Qin ZS, Kumar S, Manne U, et al. UALCAN: An update to the integrated cancer data analysis platform. Neoplasia. 2022;25:18–27. doi: 10.1016/j.neo.2022.01.001. - DOI - PMC - PubMed

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