长链非编码RNA(lncRNAs)调节其发生,口腔鳞状细胞癌(OSCC)的发生和发展。我们阐明了MAGEA4-AS1在OSCC患者中的表达特征及其作为OSCC生物标志物的活性。此外,MAGEA4-AS1上调对细胞行为的影响(增殖,评估了OSCC细胞的迁移和侵袭)和内在信号机制。首先,我们使用生物信息学方法分析了癌症基因组图谱(TCGA)OSCC中的MAGEA4-AS1表达数据,并使用qPCR分析了45对OSCC组织中的MAGEA4-AS1表达数据。然后CCK-8,乙炔基脱氧尿苷,菌落形成,进行transwell和伤口愈合试验以评估细胞增殖的变化,shMAGEA4-AS1HSC3和CAL27细胞的迁移和侵袭能力。使用cDNA末端快速扩增(RACE)测定鉴定MAGEA4-AS1的RNA序列。全转录组测序用于鉴定MAGEA4-AS1受影响的基因。此外,双荧光素酶报告系统,RNA结合蛋白免疫沉淀(RIP),进行了拯救实验以阐明MAGEA4-AS1-p53-MK2信号通路的作用。作为结果,我们发现MAGEA4-AS1在OSCC组织中表达上调。我们鉴定了MAGEA4-AS1转录物的418个核苷酸长度,并且其主要位于细胞核中。MAGEA4-AS1稳定敲低削弱了增殖,OSCC细胞的迁移和侵袭能力。机械上,p53蛋白能够激活MK2基因转录。RIP分析揭示了p53和MAGEA4-AS1之间的相互作用。MAGEA4-AS1中MK2上调下调的OSCC细胞恢复了MK2和上皮-间质转化相关蛋白的表达水平。总之,MAGEA4-AS1-p53复合物与MK2启动子结合,增强MK2的转录并激活下游信号通路,从而促进OSCC细胞的增殖和转移。MAGEA4-AS1可作为OSCC患者的诊断标志物和治疗靶点。
Long non-coding RNAs (lncRNAs) regulate the occurrence, development and progression of oral squamous cell carcinoma (OSCC). We elucidated the expression features of MAGEA4-AS1 in patients with OSCC and its activity as an OSCC biomarker. Furthermore, the impact of up-regulation of MAGEA4-AS1 on the cellular behaviors (proliferation, migration and invasion) of OSCC cells and intrinsic signal mechanisms were evaluated. Firstly, we analyzed MAGEA4-AS1 expression data in The Cancer Genome Atlas (TCGA) OSCC using a bioinformatics approach and in 45 pairs of OSCC tissues using qPCR. Then CCK-8, ethynyl deoxyuridine, colony formation, transwell and wound healing assays were conducted to assess changes in the cell proliferation, migration and invasion protential of shMAGEA4-AS1 HSC3 and CAL27 cells. The RNA sequence of MAGEA4-AS1 was identified using the rapid amplification of cDNA ends (RACE) assay. And whole-transcriptome sequencing was used to identify MAGEA4-AS1 affected genes. Additionally, dual-luciferase reporter system, RNA-binding protein immunoprecipitation (RIP), and rescue experiments were performed to clarify the role of the MAGEA4-AS1-p53-MK2 signaling pathway. As results, we found MAGEA4-AS1 was up-regulated in OSCC tissues. We identified a 418 nucleotides length of the MAGEA4-AS1 transcript and it primarily located in the cell nucleus. MAGEA4-AS1 stable knockdown weakened the proliferation, migration and invasion abilities of OSCC cells. Mechanistically, p53 protein was capable to activate MK2 gene transcription. RIP assay revealed an interaction between p53 and MAGEA4-AS1. MK2 up-regulation in MAGEA4-AS1 down-regulated OSCC cells restored MK2 and epithelial-to-mesenchymal transition related proteins\' expression levels. In conclusion, MAGEA4-AS1-p53 complexes bind to MK2 promoter, enhancing the transcription of MK2 and activating the downstream signaling pathways, consequently promoting the proliferation and metastasis of OSCC cells. MAGEA4-AS1 may serve as a diagnostic marker and therapeutic target for OSCC patients.