Background

Laryngeal squamous cell carcinoma (LSCC) is the common cancer with poor prognosis. Long non-coding RNA (lncRNA) ANRIL has been proven to play an important role in many cancers. However up to now, the role of ANRIL in LSCC is still poorly understood. The present study aimed to investigate the role and underlying mechanisms of ANRIL and miR-181a in LSCC.

Methods

Expression of ANRIL, miR-181a and Snai2 in both LSCC tissues and cells was determined by qRT-PCR. CCK-8 assay, colony formation assay, flow cytometry analysis and transwell assay were conducted to detect cell proliferation, clonogenicity, apoptosis, invasion and migration, respectively. The binding between ANRIL and miR-181a, as well miR-181a and Snai2 was confirmed by dual luciferase reporter assay. Western blotting was used to determine the protein levels of Snail, Slug, E-cadherin, N-cadherin and Vimentin.

Results

ANRIL was up-regulated while miR-181a was down-regulated in LSCC tissues. ANRIL was negatively correlated with miR-181a and was positively correlated with Snai1 and Snai2. Dual luciferase reporter assay showed ANRIL could directly sponge miR-181a to counteract its suppression on Snai2, serving as a positive regulator of Snai2. Either knockdown of ANRIL or overexpression of miR-181a significantly inhibited the proliferation, clonogenicity, invasion, migration and epithelial mesenchymal transformation (EMT), as well as promoted the apoptosis of LSCC cells, and knockdown of miR-181a reversed the effects.

Conclusion

Inhibition of ANRIL could suppress cell proliferation, clonogenicity, invasion and migration, as well as enhance cell apoptosis of LSCC cells through regulation of miR-181a/Snai2 axis, indicating that ANRIL might be a promising therapeutic target during the treatment of LSCC.

中文翻译：

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长链非编码RNA ANRIL通过调控miR-181a/Snai2轴促进喉鳞状细胞癌的增殖、克隆形成、侵袭和迁移

背景

喉鳞状细胞癌（LSCC）是常见的癌症，预后较差。长链非编码RNA（lncRNA）ANRIL已被证明在许多癌症中发挥重要作用。然而迄今为止，ANRIL 在 LSCC 中的作用仍知之甚少。本研究旨在探讨 ANRIL 和 miR-181a 在 LSCC 中的作用和潜在机制。

方法

通过 qRT-PCR 测定 LSCC 组织和细胞中 ANRIL、miR-181a 和 Snai2 的表达。CCK-8实验、集落形成实验、流式细胞仪分析和Transwell实验分别检测细胞增殖、克隆形成、凋亡、侵袭和迁移。通过双荧光素酶报告基因测定证实了 ANRIL 与 miR-181a 以及 miR-181a 与 Snai2 之间的结合。Western blotting用于测定Snail、Slug、E-钙粘蛋白、N-钙粘蛋白和波形蛋白的蛋白水平。

结果

在 LSCC 组织中，ANRIL 上调，而 miR-181a 下调。ANRIL与miR-181a呈负相关，与Snai1和Snai2呈正相关。双荧光素酶报告基因检测显示 ANRIL 可以直接海绵 miR-181a 以抵消其对 Snai2 的抑制，作为 Snai2 的正调节因子。敲低ANRIL或过表达miR-181a均显着抑制LSCC细胞的增殖、克隆形成、侵袭、迁移和上皮间质转化（EMT），并促进LSCC细胞凋亡，敲低miR-181a可逆转这种作用。

结论

抑制ANRIL可以抑制细胞增殖、克隆形成、侵袭和迁移，并通过调节miR-181a/Snai2轴增强LSCC细胞的细胞凋亡，表明ANRIL可能是LSCC治疗过程中一个有前景的治疗靶点。