RBM8A coordinately regulates expression and alternative splicing of cell cycle associated genes in AGS cells

RBM8A was knocked down in human gastric cancer cell AGS, and the transcriptome data (RNA-seq) affected by RBM8A was obtained using high-throughput sequencing technology, so as to analyze the potential target genes whose expression levels and alternative splicing patterns were regulated by RBM8A in AGS cells and analyze the functions of these genes, also we used qPCR technology to validate the results. Results:（1）RNA-seq data analysis showed that 4516 genes were differentially expressed in RBM8A knockdown AGS cells compared with the control group. upregulated genes are enriched in Type I interferon signaling pathway, apoptosis, innate immune response, cell adhesion, cell migration and other related pathways.

Downregulated genes are enriched in cell cycle, cell division, proliferation, apoptosis and other pathways. (2) Based on the RNA-seq data, we analyzed the alternative splicing events after RBM8A knockdown, and found that genes with differential alternative splicing were enriched in cell division, cell cycle related functional pathways. (3) We conducted an integrated analysis of DEG and alternative splicing events genes.

The results showed that the expression level and alternative splicing level of 816 genes were significantly different. These genes are enriched in cell cycle, cell division, mitotic spindle organization and other related biological processes (4)qPCR was used to verify the genes and the alternative splicing events Regulated by RBM8A, compared with the control group, the changes in the expression of CTNNA1, IFIT2, CENPW, HDAC2, MIF, SKA2, MCM7 and CEP55 genes in the experimental group were consistent with expectations, the alternative splicing events of genes like KMT5A,DMTF1,VRK1,AURKB,BECN1,CDKN3,CENPE and LIG3 as anticipated.

Conclusion: RBM8A may play an important regulatory role in GC cells by regulating cell cycle, cell division, proliferation, apoptosis, cell adhesion and migration through effecting the gene expression and alternative splicing events in GC cells. This study provides a reference for the current understanding of the mechanism of RBM8A carcinogenesis and cancer progression.