Abstract
BackgroundLong non-coding RNA (lncRNA) actin filament associated protein 1 antisense RNA1 (AFAP1-AS1) is oriented in an antisense direction to the protein-coding gene AFAP1 in the opposite strand. Previous studies showed that lncRNA AFAP1-AS1 was upregulated and acted as an oncogene in a variety of tumors. However, the expression and biological functions of lncRNA AFAP1-AS1 in tongue squamous cell carcinoma (TSCC) are still unknown.MethodsThe expression level of AFAP1-AS1 was measured in 103 pairs of human TSCC tissues and corresponding adjacent normal tongue mucous tissues. The correlation between AFAP1-AS1 and the clinicopathological features was evaluated using the chi-square test. The effects of AFAP1-AS1 on TSCC cells were determined via a CCK-8 assay, clone formation assay, flow cytometry, wound healing assay and transwell assay. Furthermore, the effect of AFAP1-AS1 knockdown on the activation of the Wnt/β-catenin signaling pathway was investigated. Finally, CAL-27 cells with AFAP1-AS1 knockdown were subcutaneously injected into nude mice to evaluate the effect of AFAP1-AS1 on tumor growth in vivo.ResultsIn this study, we found that lncRNA AFAP1-AS1 was increased in TSCC tissues and that patients with high AFAP1-AS1 expression had a shorter overall survival. Short hairpin RNA (shRNA)-mediated AFAP1-AS1 knockdown significantly decreased the proliferation of TSCC cells. Furthermore, AFAP1-AS1 silencing partly inhibited cell migration and invasion. Inhibition of AFAP1-AS1 decreased the activity of the Wnt/β-catenin pathway and suppressed the expression of EMT-related genes (SLUG, SNAIL1, VIM, CADN, ZEB1, ZEB2, SMAD2 and TWIST1) in TSCC cells. In addition, CAL-27 cells with AFAP1-AS1 knockdown were injected into nude mice to investigate the effect of AFAP1-AS1 on tumorigenesis in vivo. Downregulation of AFAP1-AS1 suppressed tumor growth and inhibited the expression of EMT-related genes (SLUG, SNIAL1, VIM, ZEB1, NANOG, SMAD2, NESTIN and SOX2) in vivo.ConclusionsTaken together, our findings present a road map for targeting the newly identified lncRNA AFAP1-AS1 to suppress TSCC progression, and these results elucidate a novel potential therapeutic strategy for TSCC.
Highlights
Background unknown and must be investigatedIn this study, weHead and neck cancer is the sixth most common malig- sought to determine the expression of AFAP1-AS1 in nant cancer worldwide [1,2,3], and epithelial tumors ari- tongue squamous cell carcinoma (TSCC) tissues and paired noncancerous tissues and the sing in the oral cavity are the most frequent tumors in relationship between the expression of AFAP1-AS1 and the head and neck region
In this study, we found that Long non-coding RNAs (lncRNAs) AFAP1-AS1 was increased in TSCC tissues and that patients with high
Short hairpin RNA-mediated AFAP1-AS1 knockdown significantly decreased the proliferation of TSCC cells
Summary
Head and neck cancer is the sixth most common malig- sought to determine the expression of AFAP1-AS1 in nant cancer worldwide [1,2,3], and epithelial tumors ari- TSCC tissues and paired noncancerous tissues and the sing in the oral cavity are the most frequent tumors in relationship between the expression of AFAP1-AS1 and the head and neck region. Further functional studies reoma (TSCC) is the most common epithelial cancer iden- vealed that knockdown of AFAP1-AS1 could result in tified in the oral cavity and accounts for approximately 25 to 40% of cases [1, 3]. Recent developments have been achieved in the therapeutic management of TSCC, such as surgery, chemotherapy and radiotherapy, the overall survival among TSCC patients with locally advanced disease and cervical lymph node metastasis remains dismal [4].
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