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Effect of OGT knockdown on gene expression in KGN ovarian granulosa cells

ABSTRACT: This study aimed to elucidate the spatiotemporal expression pattern of O-GlcNAc transferase (OGT) and its catalyzed O-GlcNAc modification in ovarian tissues, and to investigate their specific functions in ovarian granulosa cells. To this end, high-throughput RNA sequencing was performed to analyze genome-wide transcriptional changes following OGT knockdown in the human ovarian granulosa cell line KGN. Two specific siRNAs targeting OGT (siOGT1 and siOGT2) and a negative control siRNA (siNC) were designed. Each group included two biological replicates, resulting in a total of six transcriptomic libraries. Sequencing was carried out on the Illumina NovaSeq 6000 platform with paired-end 150-bp reads. By comparing the siOGT-treated groups with the siNC control group, 300 and 487 differentially expressed genes (DEGs) were identified in the siOGT1 and siOGT2 groups, respectively, with 176 DEGs overlapping between the two groups. Pathway enrichment analysis indicated that OGT knockdown may affect pathways related to immune defense response, cellular reaction to external stimuli, and cytokine-mediated signaling. This dataset provides a transcriptomic profile of ovarian granulosa cells under OGT interference and serves as a key resource for further investigation into the regulatory mechanisms of the OGT/O-GlcNAc axis in ovarian physiology and pathology.

ORGANISM(S): Homo sapiens

PROVIDER: GSE316021 | GEO | 2026/01/16

REPOSITORIES: GEO

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Project description:Drosophila development is a complex and dynamic process regulated, in part, by members of the Polycomb (Pc), Trithorax (Trx) and Compass chromatin modifier complexes. O-GlcNAc Transferase (OGT/SXC) is essential for Pc repression suggesting that the O-GlcNAcylation of proteins plays a key role in regulating development. OGT transfers N-acetyl-D-glucosamine (GlcNAc) onto hydroxyl groups of serine or threonine residues of key transcriptional regulators using the nutrient-derived UDP-GlcNAc as a substrate, which is dynamically removed by O-GlcNAcase (OGA). We performed ChIP-chip and microarray analysis after OGT or OGA RNAi knockdown in Drosophila S2 cells and found that O-GlcNAc was elevated genome wide particularly at genes related to mitosis and cell cycle in OGA RNAi cells, but not at sites co-occupied by Pc member Pleiohomeotic (Pho), such as the Hox and NK homeobox gene clusters. Microarray analysis suggested that altered O-GlcNAc cycling perturbed the expression of genes associated with morphogenesis and cell cycle regulation. To examine the in vivo consequences of disturbed O-GlcNAc cycling in the whole animal, we produced a null allele of oga (ogadel.1) in Drosophila. Epigenetic activators including Trx group members Trithorax (Trx), Absent small or homeotic discs 1 (Ash1) and Compass member Set1 histone methyltransferases are O-GlcNAc modified in ogadel.1 mutants. ogadel.1 mutants displayed altered expression of a distinct set of cell cycle related genes in ovaries. Our results suggest that the loss of OGA could affect epigenetic machinery by accumulating O-GlcNAc on numerous chromatin factors including Trx, Ash1 and Set1 in Drosophila. We performed affymetrix tilingarray analysis after OGT or OGA RNAi knockdown in Drosophila S2 cells to find if that O-GlcNAc was elevated genome wide particularly at genes related to mitosis and cell cycle in OGA RNAi cells, but not at sites co-occupied by Pc member Pleiohomeotic (Pho), ------------------------------- This represents the gene expression component only

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Effect of OGT knockdown on gene expression in KGN ovarian granulosa cells

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