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. 2021 Oct;10(3):420-428.
doi: 10.52547/rbmb.10.3.420.

Investigation of Decitabine Effects on HDAC3 and HDAC7 mRNA Expression in NALM-6 and HL-60 Cancer Cell Lines

Sina Dalvand  1 , Amin Namdari  2 , Farzad Sepahvand  2 , Mohammad Hassan Meshkibaf  2 , GholamReza Ahmadpour  3
Affiliations

Investigation of Decitabine Effects on HDAC3 and HDAC7 mRNA Expression in NALM-6 and HL-60 Cancer Cell Lines

Sina Dalvand et al. Rep Biochem Mol Biol. 2021 Oct.

Abstract

Background: Decitabine is a potent anticancer hypomethylating agent and changes the gene expression through the gene's promoter demethylation and also independently from DNA demethylation. So, the present study was designed to distinguish whether Decitabine, in addition to inhibitory effects on DNA methyltransferase, can change HDAC3 and HDAC7 mRNA expression in NALM-6 and HL-60 cancer cell lines.

Methods: HL-60, NALM-6, and normal cells were cultured, and the Decitabine treatment dose was obtained (1 µM) through the MTT assay. Finally, HDAC3 and HDAC7 mRNA expression were measured by Real-Time PCR in HL-60 and NALM-6 cancerous cells before and after treatment. Furthermore, HDAC3 and HDAC7 mRNA expression in untreated HL-60 and NALM-6 cancerous cells were compared to normal cells.

Results: Our results revealed that the expression of HDAC3 and HDAC7 in HL-60 and NALM-6 cells increases as compared to normal cells. After treatment of HL-60 and NALM-6 cells with Decitabine, HDAC3, and HDAC7 mRNA expression were decreased significantly.

Conclusion: Our data confirmed that the effects of Decitabine are not limited to direct hypomethylation of DNMTs, but it can indirectly affect other epigenetic factors, such as HDACs activity, through converging pathways.

Keywords: Decitabine; HDAC3; HDAC7; HL-60; NALM-6.

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Figures

Fig. 1
Fig. 1
Cell viability and IC50. A) MTT-assay diagram at different concentrations of decitabine (from 0.05 to 100 µM) for 24 h, 48 h, and 72 h. Cell viability decreased significantly only at concentrations of higher than 1 µM in comparison to control-treated cells.
Fig. 2
Fig. 2
HDAC3 expression in NALM-6, Hl-60, and normal control groups (B-cell precursor normal cell line for comparison with NALM-6 and human promyelocytic normal cell line for comparison with Hl-60). A) The expression of HDAC3 gene in the Hl-60 cancer cell line and the healthy control group (as calibrator) after normalization to GAPDH. B) the expression of HDAC3 gene in the NALM-6 cancer cell line and the healthy control group (as calibrator) after normalization to GAPDH. After analyzing with one-way ANOVA, the values were considered significant with p< 0.05 (**).
Fig. 3
Fig. 3
The chart of HDAC7 gene expression in NALM-6, Hl-60, and normal control groups (B-cell precursor Normal cell line for comparison with NALM-6 and human promyelocytic normal cell line for comparison with Hl-60). A) The expression level of the HDAC7 gene in the Hl-60 cancer cell line and the healthy control group (as calibrator) after normalization to GAPDH. B) The expression of the HDAC7 gene in the NALM-6 cancer cell line and the healthy control group (as calibrator) after normalization to GAPDH. After analyzing with one-way ANOVA, the values were considered significant with p< 0.05 (**).
Fig. 4
Fig. 4
Real-Time PCR gel patterns (qPCR) of HDAC3 and HDAC7, under non-saturating conditions on a 3% Agarose gel and stained with ethidium bromide. A) HDAC3 genes expression in an untreated HL-60 cell line (111 bp), B) HDAC3 gene expression in an untreated NALM-6 cell line (111 bp). C) HDAC3 gene expression in a treated HL-60 cell line with 1 µM Decitabine (111 bp), D) HDAC3 gene expression in a treated NALM-6 cell line with 1 µM Decitabine (111 bp), E) HDAC7 gene expression in an untreated HL-60 cell line (125 bp), F) HDAC7 gene expression in an untreated NALM-6 cell line (125 bp), G) HDAC7 genes expression in a treated HL-60 cell line with 1 µM Decitabine (125 bp), H) HDAC7 gene expression in treated NALM-6cell line with 1 µM Decitabine (125 bp).
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