上海口腔医学

上海口腔医学 ›› 2020, Vol. 29 ›› Issue (3): 267-274.doi: 10.19439/j.sjos.2020.03.008

• 论著 • 上一篇    下一篇

LncRNA NEAT1通过miR-339-5p/ITGA3轴调控舌鳞状细胞癌增殖、迁移及侵袭的分子机制

李羽1, 黄文青2, 陈林林1   

  1. 1.江西省口腔生物医学重点实验室,南昌大学附属口腔医院 口腔颌面外科, 2.牙体牙髓病科,江西 南昌 330006
  • 收稿日期:2019-08-23 修回日期:2019-10-16 出版日期:2020-06-25 发布日期:2020-07-29
  • 通讯作者: 陈林林,E-mail：oral_surgery@sina.com
  • 作者简介:李羽（1984-）,男,硕士,主治医师,E-mail：22440434@qq.com

LncRNA NEAT1 regulates proliferation, migration and invasion of tongue squamous cell carcinoma cells by regulating miR-339-5p/ITGA3 axis

LI Yu1, HUANG Wen-qing2, CHEN Lin-lin1   

  1. 1. Department of Oral and Maxillofacial Surgery, 2. Department of Endodontics, Key Laboratory of Oral Biomedicine in Jiangxi Province, Affiliated Stomatological Hospital, Nanchang University. Nanchang 330006, Jiangxi Province, China
  • Received:2019-08-23 Revised:2019-10-16 Online:2020-06-25 Published:2020-07-29

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摘要： 目的 探讨LncRNA NEAT1通过miR-339-5p/ITGA3轴调控舌鳞状细胞癌细胞增殖、迁移及侵袭的分子机制。方法 采用qRT-PCR和Western免疫印迹检测25例人舌鳞状细胞癌组织、与其对应的癌旁组织、人正常口腔黏膜细胞株HOK和人舌鳞状细胞癌细胞株Tscca、CAL27、SCC15、HN13中NEAT1、miR-339-5p、ITGA3 mRNA和ITGA3蛋白的表达。分别构建抑制NEAT1和过表达miR-339-5p的CAL27细胞株。利用MTT法检测细胞活力,Transwell法检测细胞的迁移和侵袭能力,Western免疫印迹检测CyclinD1和MMP-9蛋白的表达。采用双荧光素酶报告基因验证NEAT1、miR-339-5p和ITGA3的靶向关系,通过Western免疫印迹和qRT-PCR检测其调控关系。采用SPSS 17.0软件包对数据进行统计学分析。结果 与人正常口腔黏膜细胞株HOK相比,人舌鳞状细胞癌细胞株中NEAT1和ITGA3表达上调,miR-339-5p表达下调。抑制NEAT1或过表达miR-339-5p均可显著抑制CAL27细胞的增殖、迁移和侵袭,显著抑制CyclinD1和MMP-9蛋白的表达。双荧光素酶报告基因证实NEAT1靶向作用miR-339-5p并下调其表达水平,miR-339-5p可靶向负调控ITGA3的表达。抑制NEAT1可逆转抑制miR-339-5p表达对CAL27细胞增殖、迁移及侵袭的抑制作用。结论 LncRNA NEAT1通过下调miR-339-5p/ITGA3轴,进而促进舌鳞状细胞癌细胞增殖、迁移及侵袭。

关键词: 舌鳞状细胞癌, LncRNA NEAT1, miR-339-5p, ITGA3

Abstract: PURPOSE: To investigate the molecular mechanism of LncRNA NEAT1 regulating proliferation, migration and invasion of tongue squamous cell carcinoma cells by regulating miR-339-5p/ITGA3 axis. METHODS: qRT-PCR and Western blot were used to detect the expression of NEAT1, miR-339-5p, ITGA3 mRNA and ITGA3 protein in 25 cases of human tongue squamous cell carcinoma, its corresponding adjacent tissues, human normal oral mucosal cell line HOK and human tongue squamous cell carcinoma cell lines TSCCA, CAL27, SCC15 and HN13. CAL27 cell lines that inhibited NEAT1 and overexpressed miR-339-5p were constructed, respectively. Cell viability was detected by MTT assay, cell numbers of migration and invasion were detected by Transwell assay, and the expression of Cyclin D1 and MMP-9 proteins were detected by Western blotting. The dual luciferase reporter gene was used to verify the targeting relationship of NEAT1, miR-339-5p and ITGA3, and the regulatory relationship was detected by Western blotting and qRT-PCR. SPSS 17.0 software package was used for statistical analysis of the data. RESULTS: Compared with normal human oral mucosal cell line HOK, the expression of NEAT1 and ITGA3 was up-regulated, while the expression of miR-339-5p was down-regulated in human tongue squamous cell carcinoma cell lines. Inhibition of NEAT1 or over-expression of miR-339-5p significantly inhibited proliferation, migration and invasion of CAL27 cells, and significantly inhibited expression of Cyclin D1 and MMP-9 proteins. Dual luciferase reporter gene assay confirmed that NEAT1 directly interacted with miR-339-5p and suppressed its expression. miR-339-5p negatively regulated ITGA3 expression. Inhibition of NEAT1 reversed the inhibitory effect of the inhibition of miR-339-5p on proliferation, migration and invasion of CAL27 cells. CONCLUSIONS: LncRNA NEAT1 promotes proliferation, migration and invasion of tongue squamous cell carcinoma cells by down-regulating miR-339-5p/ITGA3 axis.

Key words: Tongue squamous cell carcinoma, LncRNA NEAT1, miR-339-5p, ITGA3

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