miR-7704对头颈鳞状细胞癌细胞生物学行为的影响

›› 2019, Vol. 10 ›› Issue (4): 196-200.

miR-7704对头颈鳞状细胞癌细胞生物学行为的影响

王春笛¹,²,郑旸³,董一博³,宋晓萌³,吴煜农³

1. 南京医科大学附属口腔医院
2. 南京医科大学口腔疾病研究江苏省重点实验室
3. 南京医科大学附属江苏省口腔医院

收稿日期:2019-08-16 修回日期:2019-10-18 出版日期:2019-12-25 发布日期:2020-01-06
通讯作者: 吴煜农 E-mail:yunongwu@aliyuan.com
基金资助:
国家自然科学基金;江苏省医学创新团队资助项目;江苏省高校优势学科建设工程

The effects of miR-7704 on the biological behavior of HNSCC cells

Received:2019-08-16 Revised:2019-10-18 Online:2019-12-25 Published:2020-01-06

摘要/Abstract

摘要： [摘要] 目的：探究miR-7704表达水平对头颈鳞状细胞癌（HNSCC）细胞生物学行为的影响。方法：应用实时定量逆转录聚合酶链式反应（qRT-PCR）检测人正常口腔上皮细胞系HOK和HNSCC细胞系HN6、HN13、HN4及CAL27中miR-7704表达水平差异。构建过表达载体miR-7704 mimic和mimic NC转染HN6、CAL27，qRT-PCR验证miR-7704表达水平上升。使用CCK8、平板克隆形成、创伤愈合实验及Transwell侵袭实验等方法探究过表达miR-7704对HNSCC细胞增殖、迁移和侵袭等生物学行为的影响。同时提取细胞总蛋白，Western blot实验检测miR-7704表达上调后上皮-间充质转化（EMT）相关指标及苏氨酸激酶（AKT）信号通路各组分表达变化情况。结果：相较HOK细胞系，HNSCC各细胞系内miR-7704表达量均明显上调。HNSCC细胞转染过表达载体miR-7704 mimic后，miR-7704表达水平上升，细胞增殖、迁移及侵袭能力增强。Western blot实验结果显示过表达miR-7704后HNSCC细胞中上皮性钙黏附蛋白（E-cadherin）及β-连环蛋白（β-catenin）表达水平下降，神经性钙黏附蛋白（N-cadherin）、波形蛋白（vimentin）、Snail蛋白及Slug蛋白表达水平均上调；苏氨酸激酶（AKT）及磷酸化苏氨酸激酶（p-AKT）蛋白表达水平均明显升高。结论：miR-7704可能作为一种促癌因子，通过调控AKT信号通路诱导EMT从而促进HNSCC细胞生物学行为进程。

关键词: miR-7704, 头颈鳞状细胞癌, 上皮-间充质转化, AKT信号通路

Abstract: [Abstract] Objective: To investigate the influence of miR-7704 expression on the biological behavior of head and neck squamous cell carcinoma (HNSCC) cells. Methods: The discrepant expression levels of miR-7704 were detected by real-time quantitative RT-PCR (qRT-PCR) among human oral epithelial cell line HOK and HNSCC cell lines, HN6, HN13, HN4 and CAL27. The expression level of miR-7704 was confirmed by qRT-PCR when HN6 and CAL27 was transfected with miR-7704 mimic and mimic NC. And, the effects of miR-7704 overexpression on the proliferation, migration and invasion of HNSCC cells were investigated by using CCK8, plate colony formation, wound healing experiment and Transwell invasion experiment. At the same time, total cell proteins were extracted, and the expression levels of EMT related indicators and threonine kinase (AKT) signaling pathway were detected by Western blot after the increase of miR-7704 expression. Results: Compared with HOK cell line, the expression levels of miR-7704 in all HNSCC cell lines were significantly up-regulated. After transfection of HNSCC cells with an overexpressed plasmid miR-7704 mimic, the expression level of miR-7704 was increased. Meanwhile, the cell proliferation, migration and invasion ability were enhanced. Western blot results showed that the expression levels of e-cadherin and beta-catenin in HNSCC cells were decreased after the overexpression of miR-7704, when the up-regulated expression of vimentin, Snail and Slug protein was observed. Levels of threonine kinase (AKT) and phosphorylated threonine kinase (p-akt) were significantly elevated. Conclusion: miR-7704 may act as a pro-oncogenic factor and induce EMT by regulating the AKT signaling pathway to promote the biological behavior of HNSCC cells.

Key words: miR-7704, Head and neck squamous cell carcinoma, Epithelial-mesenchymal transformation, AKT signaling pathway

王春笛郑旸董一博宋晓萌吴煜农. miR-7704对头颈鳞状细胞癌细胞生物学行为的影响[J]. , 2019, 10(4): 196-200.

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