背景:肝细胞癌(HCC)是全球癌症相关死亡的主要原因,受异常circRNA表达的影响。研究circRNA-miRNA-mRNA相互作用可以揭示HCC的潜在机制并确定潜在的治疗靶标。
方法:在本研究中,我们对mRNA进行了差异分析,miRNA,和circRNAs,并使用诸如miRanda之类的各种数据库建立了他们的关系,miRDB,还有mitarBase.此外,进行了功能富集和免疫浸润分析,以评估关键基因的作用。我们还进行了qPCR分析和蛋白质印迹(WB)来检查circRNA的表达水平,HCC细胞和临床样品中的CCL25和MAP2K1。此外,我们利用circ_0000069的过表达和敲低技术进行伤口愈合,transwell入侵测定,以及评估肝癌细胞迁移和侵袭能力的肿瘤发生实验。
结果:我们的发现揭示了与正常肝组织相比,HCC样品中612个上调基因和1173个下调基因的显著差异表达。此外,鉴定了429个上调的circRNAs和453个下调的circRNAs。重要的是,circ_0000069在HCC组织和细胞系中表现出上调。circ_0000069的过表达显著增加了Huh7细胞的侵袭和迁移能力,而circ_0000069的下调降低了HepG2细胞中的这种能力。此外,CCL25沉默或过表达抵消了这种影响,分开。动物研究进一步证实,hsa_circ_0000069的过表达促进异种移植裸鼠的肿瘤生长,而CCL25的抑制减弱了这种作用。
结论:Circ_0000069似乎通过调节CCL25促进HCC进展,表明circ_0000069和CCL25均可作为潜在的治疗靶点。
BACKGROUND: Hepatocellular carcinoma (HCC) is a leading cause of cancer-related deaths globally, influenced by aberrant circRNA expression. Investigating circRNA-miRNA-mRNA interactions can unveil underlying mechanisms of HCC and identify potential therapeutic targets.
METHODS: In this study, we conducted differential analyses of mRNAs, miRNAs, and circRNAs, and established their relationships using various databases such as miRanda, miRDB, and miTarBase. Additionally, functional enrichment and immune infiltration analyses were performed to evaluate the roles of key genes. We also conducted qPCR assays and western blotting (WB) to examine the expression levels of circRNA, CCL25, and MAP2K1 in both HCC cells and clinical samples. Furthermore, we utilized overexpression and knockdown techniques for circ_0000069 and conducted wound healing, transwell invasion assays, and a tumorigenesis experiment to assess the migratory and invasive abilities of HCC cells.
RESULTS: Our findings revealed significant differential expression of 612 upregulated genes and 1173 downregulated genes in HCC samples compared to normal liver tissue. Additionally, 429 upregulated circRNAs and 453 downregulated circRNAs were identified. Significantly, circ_0000069 exhibited upregulation in HCC tissues and cell lines. The overexpression of circ_0000069 notably increased the invasion and migration capacity of Huh7 cells, whereas the downregulation of circ_0000069 reduced this capability in HepG2 cells. Furthermore, this effect was counteracted by CCL25 silencing or overexpression, separately. Animal studies further confirmed that the overexpression of hsa_circ_0000069 facilitated tumor growth in xenografted nude mice, while the inhibition of CCL25 attenuated this effect.
CONCLUSIONS: Circ_0000069 appears to promote HCC progression by regulating CCL25, suggesting that both circ_0000069 and CCL25 can serve as potential therapeutic targets.