Wang Jian-Xing, Liu Yan, Jia Xin-Ju, Liu Shu-Xia, Dong Jin-Hui, Ren Xiu-Min, Xu Ou, Zhang Hai-Zhong, Duan Hui-Jun, Shan Chun-Guang
1Department of Pathology, Hebei Medical University, 361 Zhongshan East Road, Shijiazhuang, 050017 People's Republic of China.
2Department of Otolaryngology, The Second Hospital of Hebei Medical University, 215 Heping West Road, Shijiazhuang, 050000 People's Republic of China.
Cancer Cell Int. 2019 Jul 29;19:196. doi: 10.1186/s12935-019-0924-9. eCollection 2019.
Accumulating evidence shows that circular RNAs (circRNAs) plays vital roles in tumor progression. However, the biological functions of circRNAs in laryngeal squamous cell carcinoma (LSCC) metastasis is still unclear.
qRT-PCR was used to detect circFLNA, miRNAs and FLNA mRNA expression. Transwell assay and western blot were performed to evaluate cell migration ability and to detect FLNA, MMP2 and MLK1 protein expression, respectively. RNA pull-down analysis was used to find the binding-miRNAs of circFLNA. Luciferase reporter assay was used to examine the effect of circFLNA on miRNAs and miR-486-3p on FLNA expression.
In this study, we confirmed that a Filamin A (FLNA)-derived hsa_circ_0092012 known as circFLNA, was upregulated in LSCC, and the higher expression of circFLNA was correlated with LSCC lymph node metastasis. Increased circFLNA facilitates LSCC cell migration ability through upregulating FLNA and MMP2 protein expression. Mechanistically, we find that circFLNA sponges miR-486-3p in LSCC cells, relieving miR-486-3p-induced repression of FLNA which promotes LSCC cell migration. Accordingly, FLNA mRNA is overexpressed in LSCC tissues and a higher FLNA level is correlated with poor survival. Dysregulation of the circFLNA/miR-486-3p/FLNA regulatory pathway contributes to LSCC migration.
In summary, our study sheds light on the regulatory mechanism of circFLNA in LSCC migration via sponging miR-486-3p, which downregulates the FLNA protein expression. Targeting circFLNA/miR-486-3p/FLAN axis provides a potential therapeutic target for aggressive LSCC.
越来越多的证据表明，环状RNA（circRNAs）在肿瘤进展中发挥着至关重要的作用。然而，circRNAs在喉鳞状细胞癌（LSCC）转移中的生物学功能仍不清楚。
采用qRT-PCR检测circFLNA、微小RNA（miRNAs）和丝状肌动蛋白A（FLNA）信使核糖核酸（mRNA）的表达。分别进行Transwell实验和蛋白质免疫印迹法评估细胞迁移能力并检测FLNA、基质金属蛋白酶2（MMP2）和混合系蛋白激酶1（MLK1）蛋白的表达。采用RNA下拉分析寻找circFLNA的结合微小RNA。采用荧光素酶报告基因检测法检测circFLNA对微小RNA的影响以及微小RNA-486-3p（miR-486-3p）对FLNA表达的影响。
在本研究中，我们证实一种来源于细丝蛋白A（FLNA）的名为circFLNA的hsa_circ_0092012在LSCC中上调，且circFLNA的高表达与LSCC淋巴结转移相关。circFLNA的增加通过上调FLNA和MMP2蛋白表达促进LSCC细胞迁移能力。机制上，我们发现circFLNA在LSCC细胞中充当微小RNA-486-3p的海绵，减轻微小RNA-486-3p诱导的对促进LSCC细胞迁移的FLNA的抑制作用。因此，FLNA信使核糖核酸在LSCC组织中过表达，且较高的FLNA水平与较差的生存率相关。circFLNA/微小RNA-486-3p/FLNA调控通路的失调促成LSCC迁移。
总之，我们的研究揭示了circFLNA通过充当微小RNA-486-3p的海绵来下调FLNA蛋白表达从而在LSCC迁移中的调控机制。靶向circFLNA/微小RNA-486-3p/FLAN轴为侵袭性LSCC提供了一个潜在的治疗靶点。
