lncRNA-HOTTIP在低氧诱导舌鳞癌细胞上皮-间质转化过程中的作用探讨

doi:10.19438/j.cjoms.2019.01.005

中国口腔颌面外科杂志 ›› 2019, Vol. 17 ›› Issue (1): 26-31.doi: 10.19438/j.cjoms.2019.01.005

lncRNA-HOTTIP在低氧诱导舌鳞癌细胞上皮-间质转化过程中的作用探讨

林志立¹, 梁建平¹, 周斌²

1. 广州市第一人民医院口腔科,广东广州 510180;
2.中山大学附属孙逸仙纪念医院口腔颌面外科,广东广州 510120

收稿日期:2018-08-03 修回日期:2018-10-08 出版日期:2019-01-20 发布日期:2019-02-21
通讯作者: 周斌, E-mail：dophenchou@163.com
作者简介:林志立（1977-）,男,学士,主治医师,E-mail：785855836@qq.com
基金资助:
国家自然科学基金（81702695）

Hypoxia induces epithelial-to-mesenchymal transition of human tongue squamous cell carcinoma via lncRNA-HOTTIP

LIN Zhi-li¹, LIANG Jian-ping¹, ZHOU Bin²

1.Department of Stomatology, Guangzhou First People's Hospital. Guangzhou 510180;
2.Department of Oral and Maxillofacial Surgery, Sun Yat-sen Memorial Hospital Affiliated to Sun Yat-sen University. Guangzhou 510120, Guangdong Province, China

Received:2018-08-03 Revised:2018-10-08 Online:2019-01-20 Published:2019-02-21

摘要/Abstract

摘要： 目的探讨lncRNA-HOTTIP在低氧诱导人舌鳞癌细胞上皮-间质转化过程中的作用。方法体外正常氧（20% O₂）或低氧（1% O₂）状态下培养舌鳞癌SCC9、CAL27细胞48 h,实时定量PCR检测lncRNA-HOTTIP与HIF-1α的表达水平;通过小干扰RNA（Si-HIF1α）转染舌鳞癌细胞,检测HIF-1α敲低后lncRNA-HOTTIP的表达变化;进一步通过小干扰RNA（Si-HOTTIP）转染舌鳞癌细胞,验证敲低lncRNA-HOTTIP后E-cadherin、Fibronectin与Vimentin的蛋白和mRNA水平,以及细胞迁移能力的变化。采用SPSS13.0软件包对数据进行统计学分析。结果人舌鳞癌细胞SCC9、CAL27在低氧培养48 h后,lncRNA-HOTTIP与HIF-1α均显著升高;Si-HIF1α转染SCC9、CAL27并于低氧培养后,HIF-1α水平显著降低,lncRNA-HOTTIP表达下降。Si-HOTTIP转染SCC9、CAL27并在低氧培养后,lncRNA-HOTTIP水平降低,间质标志蛋白Vimentin和Fibronectin在蛋白和mRNA水平显著降低,上皮标志蛋白E-cadherin表达升高,细胞迁移能力显著降低。结论低氧可通过上调HIF-1α并激活lncRNA-HOTTIP表达,进而促进人舌鳞癌细胞发生上皮-间质转化及转移。

关键词: 低氧, 舌鳞状细胞癌, 上皮-间质转化, HOXA末端转录本反义RNA, 低氧诱导因子1α

Abstract: PURPOSE: To investigate the effect of lncRNA-HOTTIP on hypoxia induced epithelial-to-mesenchymal transition in human tongue squamous cell carcinoma (TSCC). METHODS: TSCC cell lines SCC9 and CAL27 were cultured under hypoxia (1% O₂) or normoxia (20% O₂) for 48 h. qRT-PCR was used to detect expressions of lncRNA-HOTTIP and HIF-1α at mRNA level. After transfection of HIF-1α with siRNA, HIF-1α and lncRNA-HOTTIP level were furthered detected. TSCC cell lines were further transfected lncRNA-HOTTIP with siRNA under hypoxia, the expression of HIF-1α, Vimentin, Fibronectin and E-cadherin, as well as invasion, were detected. The date was statistically analyzed with SPSS13.0 software package. RESULTS: Both SCC9 and CAL27 cells under hypoxia for 48 h showed increased lncRNA-HOTTIP and HIF-1α expression.After knocking down of HIF-1α with siRNA, lncRNA-HOTTIP level was significantly reduced. After knocking down of lncRNA-HOTTIP with siRNA, lncRNA-HOTTIP, Vimentin and Fibronectin were significantly downregulated but E-cadherin was upregulated under hypoxia; the invasion ability was decreased in SCC9 and CAL27 cells. CONCLUSIONS: Hypoxia increases HIF-1α and lncRNA-HOTTIP expression, and hypoxia induces epithelial-to-mesenchymal transition and metastasis of TSCC via upregulating lncRNA-HOTTIP.

Key words: Hypoxia, Tongue squamous cell carcinoma, Epithelial-to-mesenchymal transition, HOTTIP, HIF-1α

中图分类号:

R739.8

林志立, 梁建平, 周斌. lncRNA-HOTTIP在低氧诱导舌鳞癌细胞上皮-间质转化过程中的作用探讨[J]. 中国口腔颌面外科杂志, 2019, 17(1): 26-31.

LIN Zhi-li, LIANG Jian-ping, ZHOU Bin. Hypoxia induces epithelial-to-mesenchymal transition of human tongue squamous cell carcinoma via lncRNA-HOTTIP[J]. China J Oral Maxillofac Surg, 2019, 17(1): 26-31.

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lncRNA-HOTTIP在低氧诱导舌鳞癌细胞上皮-间质转化过程中的作用探讨

Hypoxia induces epithelial-to-mesenchymal transition of human tongue squamous cell carcinoma via lncRNA-HOTTIP

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