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Effect of KHSRP acetylation on gene expression in LNCaP cells

ABSTRACT: We established LNCaP cells in wihch KHSRP was knocked down along with LNCaP cells with silenced KHSRP and re-introduced HA-tagged KHSRP-WT/K205R.RNA-seq was carried out in the stable LNCaP cell lines to determine the underlying mechanism of KHSRP acetylation in PCa tumorigenesis. We then performed gene expression profiling analysis using data obtained from RNA-seq of 4 different cells.

ORGANISM(S): Homo sapiens

PROVIDER: GSE206486 | GEO | 2024/05/27

REPOSITORIES: GEO

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Project description:Breast cancer has become the most common type of cancers worldwide. Its high prevalence and malignant features are associated with various environmental factors and molecules. The KH-type splicing regulatory protein (KHSRP) participates in the development of breast cancer, while the underlying mechanisms are largely unknown. In this study, we silenced KHSRP expression in MDA-MB-231 cells by small interfering RNA (siKHSRP), and then assessed its effects on cellular features. Finally, we performed whole transcriptome sequencing (RNA-seq) experiments to explore the downstream targets of KHSRP, and validated their changed pattern using quantitative polymerase chain reaction. We found KHSRP showed higher expression level and was associated with worse prognosis in breast cancer patients. In siKHSRP samples, the proliferation, invasion, and migration abilities were significantly repressed compared with negative control (NC) samples, while the apoptosis level was increased. By investigating the RNA-seq data, we found KHSRP globally regulates the expression and alternative splicing profiles of MDA-MB-231 cells by identifying 1632 differentially expressed genes (DEGs) and 1630 HKSRP-regulated AS events (RASEs). Functional enriched analysis of DEGs demonstrated that cilium assembly and movement and extracellular matrix organization pathways were specifically enriched in up DEGs, consistent with the repressed migration and invasion abilities in siKHSRP cells. Interestingly, the cell cycle and DNA damage and repair associated pathways were enriched in both down DEGs and RASE genes, suggesting that KHSRP may modulate cell proliferation by regulating genes in these pathways. Finally, we validated the changed expression and AS patterns of genes in cell cycle and DNA damage/repair pathways. Expression levels of BIRC5, CCNA2, CDK1, FEN1, FOXM1, PTTG1, and UHRF1 were downregulated in siKHSRP samples. The AS patterns of PARK7, UBE2A, UBE2D3, NPM1, ERCC1, CSNK1E, CENPX, and SWI5 were also dysregulated in siKHSRP samples. In summary, our study comprehensively explored the downstream targets and their functions of KHSRP in breast cancer cells, highlighting the molecular mechanisms of KHSRP on the oncogenic features of breast cancer. The identified molecular targets could be served as potential therapeutic targets for breast cancer in future.

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Effect of KHSRP acetylation on gene expression in LNCaP cells

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