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Transcriptomic effects of EZH2 inhibition by DZNep in VEGF-stimulated HUVECs

ABSTRACT: Purpose: To compare transcriptome profiles of VEGF-treated HUVECs with or without the EZH2 inhibitor DZNep. Methods: HUVECs were treated with VEGF(20 ng/mL) or VEGF(20 ng/mL)+DZNep (10 μM) for 48 h. Total RNA was extracted using TRIzol, and poly(A)+ RNA was used for library construction following standard Illumina protocols. Sequencing was performed on an Illumina NovaSeq 6000 (PE150). Clean reads were obtained using fastp, mapped to the Homo sapiens genome (GRCh38) with HISAT2, assembled and quantified by StringTie, and differential expression was analyzed using edgeR. GO/KEGG analyses were performed using DAVID. Results: DZNep altered VEGF-induced gene expression, reducing pro-angiogenic genes and modulating pathways related to angiogenesis and endothelial function. Conclusions: This dataset provides RNA-seq profiling of VEGF-stimulated HUVECs with or without EZH2 inhibition and supports analysis of EZH2-regulated angiogenic signaling.

ORGANISM(S): Homo sapiens

PROVIDER: GSE311954 | GEO | 2025/12/31

REPOSITORIES: GEO

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Project description:The 3-Deazaneplanocin A (DZNep), one of S-adenosylhomocysteine (AdoHcy) hydrolase inhibitors, has shown antitumor activities in a broad range of solid tumors and acute myeloid leukemia. Here, we examined its effects on multiple myeloma (MM) cells and found that, at 500 nM, it potently inhibited growth and induced apoptosis in 2 of 8 MM cell lines. RNA from un-treated and DZNep treated cells was profiled by Affymetrix HG-U133 Plus 2.0 microarray and genes with a significant change in gene expression were determined by significance analysis of microarray (SAM) testing. ALOX5 was the most down-regulated gene (5.8-fold) in sensitive cells and was expressed at low level in resistant cells. The results were corroborated by quantitative RT-PCR. Western-blot analysis indicated ALOX5 was highly expressed only in sensitive cell line H929 and greatly decreased upon DZNep treatment. Ectopic expression of ALOX5 reduced sensitivity to DZNep in H929 cells. Furthermore, down-regulation of ALOX5 by RNA interference could also induce apoptosis in H929. Gene expression analysis on MM patient dataset indicated ALOX5 expression was significantly higher in MM patients compared to normal plasma cells. We also found that Bcl-2 was overexpressed in DZNep insensitive cells, and cotreatment with DZNep and ABT-737, a Bcl-2 family inhibitor, synergistically inhibited growth and induced apoptosis of DZNep insensitive MM cells. Taken together, this study shows one of mechanisms of the DZNep efficacy on MM correlates with its ability to down-regulate the ALOX5 levels. In addition, DZNep insensitivity might be associated with overexpression of Bcl-2, and the combination of ABT-737 and DZNep could synergistically induced apoptosis. These results suggest that DZNep may be exploited therapeutically for a subset of MM. Cells were treated with 0.5 μmol/L DZNep for 48 h. Total RNA was extracted by using the Qiagen RNeasy Mini kit (Germany). Gene expression was performed using the GeneChip® Human Genome U133A Array (Affymetrix) following the manufacturer’s instructions. Data analysis was performed using GeneSpring software from Agilent Technologies.

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Transcriptomic effects of EZH2 inhibition by DZNep in VEGF-stimulated HUVECs

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