Department of Otolaryngology-Head and Neck Surgery, The Central Hospital of Wuhan, Tongji Medical College Huazhong University of Science and Technology, Wuhan, Hubei, China (mainland).
Med Sci Monit. 2019 Jul 26;25:5552-5560. doi: 10.12659/MSM.917088.
BACKGROUND The clinical significance and biological function of long noncoding RNA SNHG12 have not been identified in laryngeal squamous cell carcinoma (LSCC). MATERIAL AND METHODS Expression levels of SNHG12, miR-129-5p, and WWP1 in LSCC tissues or cells were tested by RT-qPCR. MTT assay, flow cytometry, and Transwell assay were used to identify the progression of LSCC cells in vitro. Luciferase reporter assay was used to assess the associations among SNHG12, WWP1, and miR-129-5p. RESULTS SNHG12 was significantly overexpressed in LSCC tissues compared with adjacent normal tissues. The expression level of SNHG12 was significantly associated with T classification, lymph node metastasis, and cancer stage of LSCC. High expression of SNHG12 predicted shorter disease-free survival. Suppressing SNHG12 using siRNA inhibited proliferation and invasion and promoted apoptosis in the AMC-HN-8 LSCC cell line. SNHG12, mainly located in cytoplasm of AMC-HN-8 cells, was validated by dual luciferase reporter test and RT-qPCR to directly interact with miR-129-5p. Inhibition of miR-129-5p significantly increased proliferation and invasion of AMC-HN-8 cells and ameliorated the suppressive effects of si-SNHG12. Luciferase assay showed that miR-129-5p was able to combine with the 3'UTR region of WWP1, which is generally regarded as an E3 ubiquitin protein ligase. RT-qPCR and Western blot showed that WWP1 was positively regulated by SNHG12 and negatively regulated by miR-129-5p at the mRNA level and protein level. Overexpression of WWP1 significantly increased proliferation and invasion of laryngeal cancer cells. Moreover, when SNHG12 was suppressed, rescue of WWP1 restored the proliferation and invasion abilities of AMC-HN-8 cells. CONCLUSIONS Our study demonstrated that SNHG12 promoted LSCC cells progression via sponging miR-129-5p and potentiating WWP1 expression.
长链非编码 RNA SNHG12 在喉鳞状细胞癌（LSCC）中的临床意义和生物学功能尚未确定。
通过 RT-qPCR 检测 LSCC 组织或细胞中 SNHG12、miR-129-5p 和 WWP1 的表达水平。MTT 测定、流式细胞术和 Transwell 测定用于体外鉴定 LSCC 细胞的进展。荧光素酶报告基因测定用于评估 SNHG12、WWP1 和 miR-129-5p 之间的关联。
SNHG12 在 LSCC 组织中的表达明显高于相邻正常组织。SNHG12 的表达水平与 LSCC 的 T 分类、淋巴结转移和癌症分期显著相关。SNHG12 高表达预示着无病生存期较短。用 siRNA 抑制 SNHG12 可抑制 AMC-HN-8 LSCC 细胞系的增殖和侵袭，促进细胞凋亡。通过双荧光素酶报告基因试验和 RT-qPCR 验证，SNHG12 主要位于 AMC-HN-8 细胞的细胞质中，可直接与 miR-129-5p 相互作用。抑制 miR-129-5p 可显著增加 AMC-HN-8 细胞的增殖和侵袭，并改善 si-SNHG12 的抑制作用。荧光素酶测定表明，miR-129-5p 能够与 WWP1 的 3'UTR 区域结合，而 WWP1 通常被认为是一种 E3 泛素蛋白连接酶。RT-qPCR 和 Western blot 显示，在 mRNA 水平和蛋白水平上，WWP1 受 SNHG12 的正调控，受 miR-129-5p 的负调控。WWP1 的过表达可显著增加喉癌细胞的增殖和侵袭。此外，当 SNHG12 被抑制时，WWP1 的恢复挽救了 AMC-HN-8 细胞的增殖和侵袭能力。
本研究表明，SNHG12 通过海绵吸附 miR-129-5p 和增强 WWP1 的表达促进 LSCC 细胞的进展。
