Chen Xudong, Zhang Litao, Tang Shixiong
Department of Otolaryngology, The First Hospital of Ningbo, Ningbo, People's Republic of China.
Department of Otolaryngology, The First Hospital of Ningbo, Ningbo, People's Republic of China.
Auris Nasus Larynx. 2019 Feb;46(1):106-113. doi: 10.1016/j.anl.2018.05.005. Epub 2018 May 26.
MicroRNAs (miRNAs) are aberrantly expressed in various tumors and play a critical role in the progression and development of tumors. However, there is little information about the role of miR-4497 in laryngeal squamous cell carcinoma (LSCC). The aim of this study is to investigate the role of miR-4497 in LSCC.
MiR-4497 expression in tumor tissues and adjacent normal tissues was measured by RT-PCR. The effects of miR-4497 on cell viability and apoptosis were evaluated by the MTT assay, Flow cytometry and caspase-3 activity assay. Western blot analysis was used to measure the expression of various proteins. Bioinformatic analysis and luciferase reporter assay were applied to investigate the relationship between miR-4497 and GBX2.
We found that miR-4497 expression was downregulated in LSCC tumor tissues and cell lines compared to the normal counterparts. Overexpression of miR-4497 inhibits the proliferation and induces apoptosis of LSCC cells accompanied by the down-regulation of anti-apoptotic Bcl-2 proteins. Mechanisms investigation revealed that GBX2 is a direct target of miR-4497. miR-4497 expression was inversely correlated with GBX2 expression in LSCC tissues. Moreover, overexpression of miR-4497 leads to the activation of ERK, JNK but not p38. Inhibition of ERK by specific inhibitor SCH772984 could interfere the apoptosis induced by overexpression of miR-4497.
Therefore, our results indicate that miR-4497 may play a suppressive role in LSCC by targeting GBX2, which offer new insights into the tumorigenesis of LSCC.
微小RNA(miRNA)在多种肿瘤中异常表达,在肿瘤的进展和发展中起关键作用。然而,关于miR-4497在喉鳞状细胞癌(LSCC)中的作用知之甚少。本研究旨在探讨miR-4497在LSCC中的作用。
采用RT-PCR检测肿瘤组织和癌旁正常组织中miR-4497的表达。通过MTT法、流式细胞术和caspase-3活性测定评估miR-4497对细胞活力和凋亡的影响。采用蛋白质印迹分析检测各种蛋白质的表达。应用生物信息学分析和荧光素酶报告基因检测来研究miR-4497与GBX2之间的关系。
我们发现与正常组织相比,LSCC肿瘤组织和细胞系中miR-4497的表达下调。miR-4497的过表达抑制LSCC细胞的增殖并诱导其凋亡,同时抗凋亡Bcl-2蛋白表达下调。机制研究表明GBX2是miR-4497的直接靶点。在LSCC组织中,miR-4497的表达与GBX2的表达呈负相关。此外,miR-4497的过表达导致ERK、JNK激活,但不导致p38激活。用特异性抑制剂SCH772984抑制ERK可干扰miR-4497过表达诱导的凋亡。
因此,我们的结果表明miR-4497可能通过靶向GBX2在LSCC中发挥抑制作用,这为LSCC的肿瘤发生提供了新的见解。