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Abstract：
RNA-binding proteins (RBPs) play critical roles in genome regulation. In this study, we explored the latent function of KPNA2, which is an essential member of the RBP family, in the regulation of alternative splicing (AS) in gastric cancer (GC). We analyzed the role of KPNA2 in regulating differential expression and AS via RNA sequencing (RNA-seq) and improved RNA immunoprecipitation sequencing (iRIP-seq). Clinical specimens were used to analyze the associations between KPNA2 expression and clinicopathological characteristics. CCK8 assays, transwell assays and wound healing assays were performed to explore the effect of KPNA2/WDR62 on GC cell progression. KPNA2 was shown to be highly expressed in GC cells and tissues and associated with lymph node metastases. KPNA2 promoted the proliferation, migration and invasion of GC cells and primarily regulated exon skipping, alternative 3\'s splice sites (A3SSs), alternative 5\' splice sites (A5SSs), and cassette exons. We further revealed that KPNA2 participated in biological processes related to cell proliferation, and the immune response in GC via the regulation of transcription. In addition, KPNA2 preferentially bound to intron regions. Notably, KPNA2 regulated the A3SS AS mode of WDR62, and upregulation of WDR62 reversed the KPNA2 downregulation-induced inhibition of GC cell proliferation, migration and invasion. Finally, we discovered that the AS of immune-related molecules could be regulated by KPNA2. Overall, our results demonstrated for the first time that KPNA2 functions as an oncogenic splicing factor in GC that regulated the AS and differential expression of GC-related genes, and KPNA2 may be a potential target for GC treatment.
摘要：
RNA结合蛋白(RBP)在基因组调控中起关键作用。在这项研究中,我们探索了KPNA2的潜在功能，KPNA2是RBP家族的重要成员，在胃癌（GC）中选择性剪接（AS）的调控。我们通过RNA测序（RNA-seq）和改进的RNA免疫沉淀测序（iRIP-seq）分析了KPNA2在调节差异表达和AS中的作用。临床标本用于分析KPNA2表达与临床病理特征之间的关联。CCK8测定，进行了transwell测定和伤口愈合测定，以探索KPNA2/WDR62对GC细胞进展的影响。KPNA2在GC细胞和组织中高表达，并与淋巴结转移有关。KPNA2促进了扩散，GC细胞的迁移和侵袭，主要调节外显子跳跃，替代3的剪接位点（A3SS），替代5个剪接位点(A5SS)，和盒式磁带外显子。我们进一步揭示了KPNA2参与了与细胞增殖相关的生物学过程。而GC中的免疫应答则经由过程转录调控。此外,KPNA2优先结合内含子区域。值得注意的是,KPNA2调节WDR62的A3SSAS模式，WDR62的上调逆转了KPNA2下调诱导的GC细胞增殖抑制，移民和入侵。最后,我们发现免疫相关分子的AS可以被KPNA2调节。总的来说,我们的研究结果首次证明，KPNA2在GC中作为致癌剪接因子起作用，调节AS和GC相关基因的差异表达，KPNA2可能是GC治疗的潜在靶标。
