背景:长链非编码RNA(lncRNA)已成为肿瘤发生和肿瘤进展的关键参与者。然而,lncRNAs在结直肠癌(CRC)中的生物学功能和潜在机制尚不清楚.
方法:通过生物信息学分析鉴定了新的lncRNAPOU6F2-AS1,通过qRT-PCR和FISH验证其在CRC患者中的表达。体外和体内实验,如BODIPY染色,油红O染色,甘油三酯(TAG)测定,和液相色谱质谱(LC-MS)随后进行CRC标本和细胞,以确定临床意义,和POU6F2-AS1的功能作用。生物素化RNA下拉,RIP,Me-RIP,ChIP,和患者来源的类器官(PDO)培养测定进行,以确认POU6F2-AS1的潜在机制。
结果:lncRNAPOU6F2-AS1在CRC中明显上调,并与CRC患者的不良临床病理特征和不良总体生存率相关。功能上,POU6F2-AS1在体外和体内促进CRC细胞的生长和脂肪生成。机械上,METTL3诱导的m6A修饰参与POU6F2-AS1的上调。此外,上调的POU6F2-AS1可以将YBX1束缚到FASN启动子以诱导转录激活,从而促进CRC细胞的生长和脂肪生成。
结论:我们的数据表明,POU6F2-AS1的上调在CRC脂肪酸代谢中起着关键作用,并可能为CRC提供新的有前景的生物标志物和治疗靶点。
Long noncoding RNAs (lncRNAs) have emerged as key players in tumorigenesis and tumour progression. However, the biological functions and potential mechanisms of lncRNAs in colorectal cancer (CRC) are unclear.
The novel lncRNA POU6F2-AS1 was identified through bioinformatics analysis, and its expression in CRC patients was verified via qRT-PCR and FISH. In vitro and in vivo experiments, such as BODIPY staining, Oil Red O staining, triglyceride (TAG) assays, and liquid chromatography mass spectrometry (LC-MS) were subsequently performed with CRC specimens and cells to determine the clinical significance, and functional roles of POU6F2-AS1. Biotinylated RNA pull-down, RIP, Me-RIP, ChIP, and patient-derived organoid (PDO) culture assays were performed to confirm the underlying mechanism of POU6F2-AS1.
The lncRNA POU6F2-AS1 is markedly upregulated in CRC and associated with adverse clinicopathological features and poor overall survival in CRC patients. Functionally, POU6F2-AS1 promotes the growth and lipogenesis of CRC cells both in vitro and in vivo. Mechanistically, METTL3-induced m6A modification is involved in the upregulation of POU6F2-AS1. Furthermore, upregulated POU6F2-AS1 could tether YBX1 to the FASN promoter to induce transcriptional activation, thus facilitating the growth and lipogenesis of CRC cells.
Our data revealed that the upregulation of POU6F2-AS1 plays a critical role in CRC fatty acid metabolism and might provide a novel promising biomarker and therapeutic target for CRC.