口腔鳞状细胞癌（OSCC）是口腔常见的恶性肿瘤，约占口腔恶性肿瘤的90%。它在全球最常见的癌症类型中排名第六，每年导致约 145,000 人死亡。人们普遍认为，非编码RNA通过竞争性调节相互作用参与癌症的发展，称为竞争性内源RNA（ceRNA）网络，其中长非编码RNA（lncRNA）作为microRNA的诱饵来调节基因表达。据报道，LncRNA FOXD2-AS1 在 OSCC 中发挥致癌作用。然而，FOXD2-AS1 介导的 ceRNA 网络尚未被研究。本研究旨在探讨FOXD2-AS1对OSCC细胞进程的影响及其潜在的ceRNA机制。通过逆转录和定量聚合酶链反应测定FOXD2-AS1在OSCC细胞中的表达。将靶向 FOXD2-AS1 的短发夹 RNA 转染至 OSCC 细胞中以沉默 FOXD2-AS1 表达。然后，进行功能丧失实验（每次测定 n = 3），分别使用集落形成、TdT 介导的 dUTP Nick-End 标记、伤口愈合和 Transwell 测定来测量细胞增殖、凋亡、迁移和侵袭。 RNA 结合关系通过 RNA 免疫沉淀和荧光素酶报告基因测定进行验证。救援实验旨在验证 FOXD2-AS1 是否通过细胞视黄酸结合蛋白 2 (CRABP2) 基因影响细胞行为。通过GraphPad Prism 6.0软件和SPSS软件进行统计。FOXD2-AS1在Cal27和SCC9细胞中显着上调（6.8和6.4倍）。 FOXD2-AS1 敲除后，OSCC 细胞增殖、迁移和侵袭受到抑制（减少约 50%），而 OSCC 细胞凋亡增强（增加两倍以上）。 FOXD2-AS1 与 miR-378 g 相互作用以改变 CRABP2 表达。 CRABP2 上调部分挽救了 (*p < 0.05, **p < 0.01, ***p < 0.001) FOXD2-AS1 缺失对 OSCC 细胞恶性特征的抑制作用。FOXD2-AS1 通过与 miR 相互作用增强 OSCC 恶性细胞行为-378 g 调节 CRABP2 表达。© 2024。作者。

Oral squamous cell cancer (OSCC) is a prevalent malignancy in oral cavity, accounting for nearly 90% of oral malignancies. It ranks sixth among the most common types of cancer worldwide and is responsible for approximately 145,000 deaths each year. It is widely accepted that noncoding RNAs participate cancer development in competitive regulatory interaction, knowing as competing endogenous RNA (ceRNA) network, whereby long non-coding RNA (lncRNA) function as decoys of microRNAs to regulate gene expression. LncRNA FOXD2-AS1 was reported to exert an oncogenic role in OSCC. Nevertheless, the ceRNA network mediated by FOXD2-AS1 was not investigated yet. This study aimed to explore the effect of FOXD2-AS1 on OSCC cell process and the underlying ceRNA mechanism.FOXD2-AS1 expression in OSCC cells were determined via reverse transcription and quantitative polymerase chain reaction. Short hairpin RNA targeting FOXD2-AS1 was transfected into OSCC cells to silence FOXD2-AS1 expression. Then, loss-of-function experiments (n = 3 each assay) were performed to measure cell proliferation, apoptosis, migration, and invasion using colony formation, TdT-mediated dUTP Nick-End Labeling, wound healing and Transwell assays, respectively. RNA binding relation was verified by RNA immunoprecipitation and luciferase reporter assays. Rescue experiments were designed to validate whether FOXD2-AS1 affects cell behavior via the gene cellular retinoic acid binding protein 2 (CRABP2). Statistics were processed by GraphPad Prism 6.0 Software and SPSS software.FOXD2-AS1 was significantly upregulated in Cal27 and SCC9 cells (6.8 and 6.4 folds). In response to FOXD2-AS1 knockout, OSCC cell proliferation, migration and invasion were suppressed (approximately 50% decrease) while OSCC cell apoptosis was enhanced (more than two-fold increase). FOXD2-AS1 interacted with miR-378 g to alter CRABP2 expression. CRABP2 upregulation partly rescued (*p < 0.05, **p < 0.01, ***p < 0.001) the inhibitory impact of FOXD2-AS1 depletion on malignant characteristics of OSCC cells.FOXD2-AS1 enhances OSCC malignant cell behaviors by interacting with miR-378 g to regulate CRABP2 expression.© 2024. The Author(s).