Department of Stomatology, The First Affiliated Hospital of Zhengzhou University , Zhengzhou , Henan , P. R. China.
The Academy of Medical Science, Zhengzhou University , Zhengzhou , Henan , P. R. China.
Cell Cycle. 2019 Nov;18(21):2902-2913. doi: 10.1080/15384101.2019.1662257. Epub 2019 Sep 4.
Oral squamous cell carcinoma (OSCC) ranks as the sixth most common carcinoma worldwide, and the third most common carcinoma in developing countries as well. Recently, the aberrant expression of lncRNA CCAT1 has been revealed to play an important role in the development of several cancers. However, its role in OSCC remains unknown. The expression levels of CCAT1 and miR-181a were determined in 15 paired primary OSCC tissues and their adjacent noncancerous tissues and cell lines with qPCR. shRNA against CCAT1 was employed to investigate the impact of CCAT1 on proliferation and metastasis. Then dual luciferase reporter and RIP assays were utilized to study the interaction between CCAT1 and miR-181a. Cells transfected with sh-CCAT1 or treated with miR-181a inhibitor were subjected to western blot to investigate the role of Wnt/β-catenin signaling in CCAT1-mediated proliferation and metastasis. Finally, the role of CCAT1 in OSCC was confirmed with tumor xenografts mice model. CCAT1 was upregulated in OSCC tissues and cell lines. Knockdown of CCAT1 inhibited the proliferation, migration and invasion of OSCC cells, while the cell apoptosis was enhanced. Luciferase and RIP assays revealed that miR-181a was a direct target of CCAT1. Inhibition of miR-181a partially reversed the efficacy of sh-CCAT1. Moreover, sh-CCAT1 inhibited OSCC tissues growth through inhibiting Wnt signaling in a miR-181a-dependent manner . lncRNA CCAT1 activated Wnt/β-catenin signaling via inhibiting miR-181a, resulting in the cell proliferation, migration and invasion of OSCC, suggesting that CCAT1 might serve as a potential target of OSCC treatment. LncRNA: long non-coding RNA; OSCC: oral squamous cell carcinoma; 3' UTR: 3' untranslated region; ANOVA: one-way analysis of variance; CDK: cyclin-dependent kinase; ceRNA: competing endogenous RNA; FBS: fetal bovine serum; HGF: human gingival fibroblasts; MAPK: mitogen-activated protein kinase; miRNA: micro RNA; ncRNA: noncoding RNAs; PBS: phosphate-buffered saline; PI3K: phosphatidylinositol 3-kinase.
口腔鳞状细胞癌（OSCC）是全球第六大常见的癌种，也是发展中国家第三大常见的癌种。最近，lncRNA CCAT1 的异常表达已被揭示在几种癌症的发展中发挥重要作用。然而，其在 OSCC 中的作用尚不清楚。采用 qPCR 测定 15 对原发性 OSCC 组织及其相邻非癌组织和细胞系中 CCAT1 和 miR-181a 的表达水平。采用 shRNA 靶向 CCAT1 研究 CCAT1 对增殖和转移的影响。然后采用双荧光素酶报告基因和 RIP 测定研究 CCAT1 与 miR-181a 之间的相互作用。用 sh-CCAT1 转染或用 miR-181a 抑制剂处理细胞，通过 Western blot 研究 Wnt/β-catenin 信号通路在 CCAT1 介导的增殖和转移中的作用。最后，通过 OSCC 荷瘤小鼠模型证实了 CCAT1 在 OSCC 中的作用。CCAT1 在 OSCC 组织和细胞系中上调。敲低 CCAT1 抑制了 OSCC 细胞的增殖、迁移和侵袭，而细胞凋亡增强。荧光素酶和 RIP 测定表明 miR-181a 是 CCAT1 的直接靶标。抑制 miR-181a 部分逆转了 sh-CCAT1 的作用。此外，sh-CCAT1 通过 miR-181a 依赖性方式抑制 Wnt 信号抑制 OSCC 组织生长。长链非编码 RNA CCAT1 通过抑制 miR-181a 激活 Wnt/β-catenin 信号通路，导致 OSCC 细胞的增殖、迁移和侵袭，提示 CCAT1 可能成为 OSCC 治疗的潜在靶点。LncRNA：长链非编码 RNA；OSCC：口腔鳞状细胞癌；3'UTR：3'非翻译区；ANOVA：单因素方差分析；CDK：周期蛋白依赖性激酶；ceRNA：竞争性内源性 RNA；FBS：胎牛血清；HGF：人牙龈成纤维细胞；MAPK：丝裂原活化蛋白激酶；miRNA：微小 RNA；ncRNA：非编码 RNA；PBS：磷酸盐缓冲液；PI3K：磷酸肌醇 3-激酶。
