MicroRNA-363 targets myosin 1B to reduce cellular migration in head and neck cancer.
MicroRNA-363 targets myosin 1B to reduce cellular migration in head and neck cancer.
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DOI：
10.1186/s12885-015-1888-3
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发表时间：
2015-11-06
期刊：
影响因子：
3.8
通讯作者：
Khan SA
中科院分区：
文献类型：
--
作者：
Chapman BV;Wald AI;Akhtar P;Munko AC;Xu J;Gibson SP;Grandis JR;Ferris RL;Khan SA
头颈部鳞状细胞癌（SCCHN）仍然是一种流行和毁灭性的疾病。最近，与高危人乳头瘤病毒（HPV）感染相关的SCCHN病例有所增加。HPV阳性和HPV阴性SCCHN的临床特征是不同的，但它们的分子特征只是最近才开始出现。微小RNA（miRNAs，miRs）是一类小的非编码RNA，在肿瘤的发生和发展中起重要作用，可能作为癌基因或肿瘤抑制因子。我们实验室先前的研究表明，与HPV阴性的SCCHN细胞系相比，HPV阳性的SCCHN细胞系中miR-363过表达，并且HPV 16-E6癌蛋白上调SCCHN细胞系中的miR-363。然而，miR-363在HPV感染背景下SCCHN中的功能作用仍有待阐明。我们分析了来自癌症基因组图谱（TCGA）的已知HPV状态的SCCHN肿瘤和我们机构的独立队列中的miR-363水平。在HPV阴性细胞系中过表达miR-363后进行细胞迁移研究。利用生物信息学工具和荧光素酶报告基因测定来证实miR-363靶向肌球蛋白1B（MYO 1B）的3 '-UTR。评估HPV阴性SCCHN细胞系中miR-363过表达后MYO 1B mRNA和蛋白质表达水平。进行MYO 1B的小干扰RNA（siRNA）敲低以评估SCCHN细胞系中MYO 1B表达降低的表型意义。发现miR-363在HPV-16阳性的SCCHN肿瘤中过表达，而在HPV阴性的SCCHN肿瘤中过表达。在HPV阴性JHU 028细胞中进行的荧光素酶报告基因测定证实，miR-363靶向MYO 1B的3 'UTR中的两个潜在结合位点之一。在4种HPV阴性SCCHN细胞系中，miR-363过表达后MYO 1B mRNA和蛋白水平降低。miR-363表达增加或MYO 1B表达的siRNA敲低降低了SCCHN细胞系的Transwell迁移，表明miR-363诱导的SCCHN细胞迁移减弱可能通过MYO 1B下调起作用。这些发现表明，miR-363的过表达减少了头颈癌中的细胞迁移，并揭示了miR-363、肌球蛋白1b和HPV阳性SCCHN之间的生物学关系。本文的在线版本（doi：10.1186/s12885-015-1888-3）包含补充材料，可供授权用户使用。
Squamous cell carcinoma of the head and neck (SCCHN) remains a prevalent and devastating disease. Recently, there has been an increase in SCCHN cases that are associated with high-risk human papillomavirus (HPV) infection. The clinical characteristics of HPV-positive and HPV-negative SCCHN are known to be different but their molecular features are only recently beginning to emerge. MicroRNAs (miRNAs, miRs) are small, non-coding RNAs that are likely to play significant roles in cancer initiation and progression where they may act as oncogenes or tumor suppressors. Previous studies in our laboratory showed that miR-363 is overexpressed in HPV-positive compared to HPV-negative SCCHN cell lines, and the HPV type 16-E6 oncoprotein upregulates miR-363 in SCCHN cell lines. However, the functional role of miR-363 in SCCHN in the context of HPV infection remains to be elucidated. We analyzed miR-363 levels in SCCHN tumors with known HPV-status from The Cancer Genome Atlas (TCGA) and an independent cohort from our institution. Cell migration studies were conducted following the overexpression of miR-363 in HPV-negative cell lines. Bioinformatic tools and a luciferase reporter assay were utilized to confirm that miR-363 targets the 3’-UTR of myosin 1B (MYO1B). MYO1B mRNA and protein expression levels were evaluated following miR-363 overexpression in HPV-negative SCCHN cell lines. Small interfering RNA (siRNA) knockdown of MYO1B was performed to assess the phenotypic implication of reduced MYO1B expression in SCCHN cell lines. MiR-363 was found to be overexpressed in HPV-16-positive compared to the HPV-negative SCCHN tumors. Luciferase reporter assays performed in HPV-negative JHU028 cells confirmed that miR-363 targets one of its two potential binding sites in the 3’UTR of MYO1B. MYO1B mRNA and protein levels were reduced upon miR-363 overexpression in four HPV-negative SCCHN cell lines. Increased miR-363 expression or siRNA knockdown of MYO1B expression reduced Transwell migration of SCCHN cell lines, indicating that the miR-363-induced migration attenuation of SCCHN cells may act through MYO1B downregulation. These findings demonstrate that the overexpression of miR-363 reduces cellular migration in head and neck cancer and reveal the biological relationship between miR-363, myosin 1b, and HPV-positive SCCHN. The online version of this article (doi:10.1186/s12885-015-1888-3) contains supplementary material, which is available to authorized users.