MiR-5189-3p Suppresses cell Proliferation, Invasion and Migration Through Targeting EIF5A2 in Laryngeal Squamous Cell Carcinoma.

A growing body of evidence suggests that miR-5189-3p plays a critical role in multiple diseases. This study aimed to investigate the function of miR-5189-3p in laryngeal squamous cell carcinoma (LSCC) and explore its underlying mechanisms. qRT-PCR was designed to determine the expression levels of miR-5189-3p and eukaryotic translation initiation factor 5A2 (EIF5A2), while CCK-8 assay was performed to measure the effects of miR-5189-3p on cell proliferation. Transwell assay was performed to evaluate cell invasion as well as migration, and wound healing assay was applied to demonstrate cell migratory ability. Target gene prediction and luciferase reporter assay were developed to screen the possible target gene of miR-5189-3p, and Western blot was designed to measure EIF5A2 protein expression. MiR-5189-3p was down-regulated in LSCC tissues and cell lines. Up-regulation of miR-5189-3p notably inhibited cell proliferation, invasion, and migration in HEP2 and FADU cells. EIF5A2 was the potential downstream gene of miR-5189-3p, and overexpression of miR-5189-3p apparently reduced EIF5A2 expression. Moreover, reintroduction of EIF5A2 rescued the tumor suppressive effects of miR-5189-3p. MiR-5189-3p functions as a tumor inhibitor in LSCC progression via directly regulating EIF5A2 and may be a potential therapeutic target for LSCC.

Lou D ，Jia Q ，Zhang H ，Wang J ，Liu L ，Liu Z ，Jia X ，Wang J ，Shan C ... -  《-》

LINC00518 affects the proliferation, invasion and migration of cutaneous malignant melanoma cells via miR-526b-3p/EIF5A2 axis.

Xu J ，Zhang F ，Lin M ，Tong Y ... -  《-》

LncRNA NEAT1 promotes proliferation, migration, and invasion of laryngeal squamous cell carcinoma cells through miR-411-3p/FZD3-mediated Wnt signaling pathway.

The lncRNA NEAT1 has been shown to promote the progression of several cancers, containing laryngeal squamous cell carcinoma (LSCC). However, the precise mechanism by which it promotes LSCC progression remains unclear. In this study, we verified the high expression of lncRNA NEAT1 in LSCC tissues and cells using RT-qPCR. Analysis of clinical data exhibited that high expression of lncRNA NEAT1 was associated with a history of smoking, worse T stage, lymph node metastasis, and later TNM stage in patients with LSCC. The promotion effect of lncRNA NEAT1 on LSCC cell proliferation, migration, invasion, and tumor growth in vivo was verified by CCK-8, plate clone formation, Transwell, and nude mouse tumorigenicity assays. Bioinformatics prediction and double luciferase reporter gene assay verified the binding of miR-411-3p to lncRNA NEAT1 and FZD3 mRNA, and inhibition of miR-411-3p reversed the inhibitory effect of lncRNA NEAT1 on FZD3 expression in LSCC cells. We also verified that lncRNA NEAT1-mediated FZD3 activation in the Wnt pathway affects LSCC development. In conclusion, we demonstrate that lncRNA NEAT1 promotes the progression of LSCC, and propose that the lncRNA NEAT1/miR-411-3p/FZD3 axis may be an effective target for LSCC therapy.

Liang J ，Fang F ，Gao X ，Shi J ，Zhao J ，Zhao Y ... -  《BMC CANCER》

Anti-cancer effect of miR-139-3p on laryngeal squamous cell carcinoma by targeting rab5a: In vitro and in vivo studies.

Laryngeal squamous cell carcinoma is the second most common head and neck squamous cell carcinoma. Nowadays, as traditional treatment methods are gradually limited, the development of new treatment methods needs to be resolved. This study aimed to investigate the role of microRNA(miR)-139-3p in laryngeal squamous cell carcinoma, and further explored the underlying mechanism. In this study, we first used quantitative real time polymerase chain reaction (qRT-PCR) to detect the level of miR-139-3p in laryngeal squamous cell carcinoma tissue. Then, TargetScan and dual luciferase reporter assay were used to explore and verify whether rab5a was a direct target of miR-139-3p. Thereafter, the expression of miR-139-3p and rab5a in laryngeal squamous cell carcinoma cell line SNU46 was changed by transfection with miR-139-3p mimic or rab5a-plasmid. Then, SNU46 cell proliferation, cell apoptosis, cell cycle, cell migration and cell invasion were determined using Cell Counting Kit-8 (CCK-8), flow cytometry, scratch assay and Transwell assay, respectively. Finally, mouse tumor formation experiments were used to test whether miR-139-3p still exerted its role in inhibiting laryngeal squamous cell carcinoma in vivo. Compared with the adjacent normal tissues, miR-139-3p significantly down-regulated in laryngeal squamous cell carcinoma tissue. It was confirmed by dual luciferase reporter experiment that rab5a was a direct target of miR-139-3p. Moreover, the up-regulation of miR-139-3p could effectively inhibit the proliferation, migration and invasion of laryngeal squamous cell carcinoma cells, and induced cell cycle arrest and apoptosis. In the molecular level study, we found that up-regulation of miR-139-3p inhibited the expression of rab5a in SNU46 cells. In addition, the protein and mRNA expression of factors related to cell migration, invasion, proliferation and apoptosis, such as integrin β1, Focal adhesion kinase (FAK), paxillin, B cell lymphoma-2 (bcl-2), nuclear factor-kappaB (NF-κB), vascular endothelial growth factor (VEGF), matrix metalloproteinase 9 (MMP9), in SNU46 cells were changed after miR-139-3p up-regulation. Consistent with the results of in vitro studies, in vivo experiments showed that miR-139-3p mimic inhibited laryngeal squamous cell carcinoma tumor growth. All the effects of miR-139-3p on laryngeal squamous cell carcinoma were reversed by rab5a over-expression. miR-139-3p could inhibit laryngeal squamous cell carcinoma by targeting rab5a both in vitro and in vivo.

Ma Y ，Gong Z ，Wang H ，Liang Y ，Huang X ，Yu G ... -  《-》

Long noncoding RNA PVT1 promotes laryngeal squamous cell carcinoma development by acting as a molecular sponge to regulate miR-519d-3p.

This study was designed to investigate the effects and mechanism of long noncoding RNA (lncRNA) PVT1 on cell migration, proliferation, and apoptosis of laryngeal squamous cell carcinoma (LSCC). We screened lncRNAs expression profiles in four pair LSCC and matched noncancerous tissues by microarray assay. The messenger RNA levels of PVT1 in tissues and cells were evaluated by quantitative real-time polymerase chain reaction analysis. StarBase website was used to predict the target miRNAs for PVT1. And the interaction between PVT1 and target miRNA-519d-3p in LSCC cells was analyzed using dual-luciferase reporter assay. MTT assay was used to investigate the cell viability. Cell counting assay was used to explore the cell proliferation. Annexin-V propidium iodide flow cytometry was used to examine the cell apoptosis, and transwell assay was used to investigate the effects of lncRNA PVT1 on cell migration. PVT1 was significantly overexpressed in human LSCC tissues and several LSCC cell lines. Upregulation of lncRNA PVT1 markedly facilitated proliferation suppressed apoptosis and promoted cell migration in LSCC cells. We further demonstrated that silencing PVT1 strikingly suppressed proliferation, promoted apoptosis, and reduced migration in LSCC cells. Further bioinformatic analysis and dual-luciferase reporter assay revealed that PVT1 could function as an oncogenic transcript partly through sponging miR-519d-3p. Besides, mechanistic investigations indicated that PVT1 could promote cell and migration through interacting with miR-519d-3p. LncRNA PVT1 is consistently overexpressed in human LSCC, and overexpression of lncRNA PVT1 contributes to the proliferation and migration of LSCC through inhibiting miR-519d-3p expression.

Zheng X ，Zhao K ，Liu T ，Liu L ，Zhou C ，Xu M ... -  《-》