长链非编码 RNA AFAP1-AS1 的上调通过 Wnt/β-catenin 信号通路影响舌鳞癌细胞的增殖、侵袭和存活。
Upregulation of the long non-coding RNA AFAP1-AS1 affects the proliferation, invasion and survival of tongue squamous cell carcinoma via the Wnt/β-catenin signaling pathway.
机构信息
Department of Laboratory Medicine, The Second Xiangya Hospital, Central South University, Changsha, Hunan, 410011, China.
Department of Ophthalmology, Xiangya Hospital, Central South University, Changsha, Hunan, 410008, China.
出版信息
Mol Cancer. 2018 Jan 8;17(1):3. doi: 10.1186/s12943-017-0752-2.
BACKGROUND
Long 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.
METHODS
The 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.
RESULTS
In 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.
CONCLUSIONS
Taken 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.
背景
长链非编码 RNA（lncRNA）肌动蛋白丝相关蛋白 1 反义 RNA1（AFAP1-AS1）与蛋白编码基因 AFAP1 在相反的链上呈反义方向排列。先前的研究表明，lncRNA AFAP1-AS1 在多种肿瘤中上调并作为癌基因发挥作用。然而，lncRNA AFAP1-AS1 在舌鳞状细胞癌（TSCC）中的表达和生物学功能仍不清楚。
方法
检测 103 对人 TSCC 组织及其相应的相邻正常舌黏膜组织中 AFAP1-AS1 的表达水平。采用卡方检验评估 AFAP1-AS1 与临床病理特征的相关性。通过 CCK-8 检测、集落形成检测、流式细胞术、划痕愈合检测和 Transwell 检测，确定 AFAP1-AS1 对 TSCC 细胞的影响。此外，研究了 AFAP1-AS1 敲低对 Wnt/β-catenin 信号通路激活的影响。最后，将 AFAP1-AS1 敲低的 CAL-27 细胞皮下注射到裸鼠体内，评估 AFAP1-AS1 对体内肿瘤生长的影响。
结果
在这项研究中，我们发现 lncRNA AFAP1-AS1 在 TSCC 组织中上调，高表达 AFAP1-AS1 的患者总生存期较短。短发夹 RNA（shRNA）介导的 AFAP1-AS1 敲低显著降低了 TSCC 细胞的增殖。此外，AFAP1-AS1 沉默部分抑制了细胞迁移和侵袭。抑制 AFAP1-AS1 降低了 Wnt/β-catenin 通路的活性，并抑制了 EMT 相关基因（SLUG、SNAIL1、VIM、CADN、ZEB1、ZEB2、SMAD2 和 TWIST1）在 TSCC 细胞中的表达。此外，将 AFAP1-AS1 敲低的 CAL-27 细胞注射到裸鼠体内，以研究 AFAP1-AS1 对体内肿瘤发生的影响。下调 AFAP1-AS1 抑制了肿瘤生长，并抑制了 EMT 相关基因（SLUG、SNAIL1、VIM、ZEB1、NANOG、SMAD2、NESTIN 和 SOX2）在体内的表达。
结论
综上所述，我们的研究结果为靶向新鉴定的 lncRNA AFAP1-AS1 抑制 TSCC 进展提供了路线图，并阐明了 TSCC 潜在的治疗新策略。
Zotero 插件