Building Of A Panel Of MiRNAs As A Prognosis Model For Oral Squamous Carcinoma And Specific Study On MiRNA-548k Promoting Tumor Progression In OSCC

Oral squamous cell carcinoma(OSCC)is the sixth most common malignancy in the world.OSCC is a common and fatal malignancy among head and neck malignant neoplasms.Therefore,there is a requirement to identify more effective treatment strategies to improve the survival rate and reduce complications of patients with OSCC.Micro RNAs(mi RNAs/mi Rs)have attracted increasing attention over the past years as their roles in malignant tumors.The Cancer Genome Atlas(TCGA)is a landmark cancer genomics program,that molecularly characterizes over 20,000 primary cancer and matches normal samples spanning 33 cancer types.A number of prediction models based on mi RNA expression were proposed for tumor survival.As one of the most differentiated mi RNAs in the prediction model of oral squamous cell carcinoma,mi RNA may play an important role in the development of OSCC.Objective:In this study,we establish a prognostic model based on a mi RNA panel to better predict the survival of Head and neck squamous cell carcinoma(HNSCC)patients.Besides,we investigate the potential role of the relevant mi RNAs in OSCC.Methods:The expression data of mi RNAs in patients with head and neck squamous cell carcinoma and relevant clinical data were obtained using TCGA database to construct a survival and prognosis model based on mi RNA expression level.mi RNA that significantly affected the prognosis of OSCC patients were selected as research subjects.q RT-PCR was used to further verify the expression level of mi RNA-548 k in the OSCC samples.mi RNA-548 k mimics and mi RNA-548 k inhibitor were used for cell function test to determine the possible ways to promote tumor progression.Potential downstream targets of mi RNA-548 k were identified by bioinformatics.The downstream was identified by the double luciferase reporter method,and si RNA were constructed for downstream functional experiments.In order to further verify the role of mi R-548 k in in vivo experiments in mice,a mouse PDX model was constructed by using human OSCC tissue,and the tumor injected with mi R-548 k was observed compared with those in the negative control group.Results:Using this data,we identified 140 differentially expressed mi RNAs(DEMs)between HNSCC tissue samples and adjacent normal tissue samples.Our prognostic model includes seven mi RNAs(i.e.hsa-mi R-499 a,hsa-mi R-548 k,hsa-mi R-3619,hsa-mi R-99 a,hsa-mi R-137,hsa-mi R-3170,and hsa-mi R-654).The outstanding performance of the seven-mi RNA prognostic model was confirmed by time-dependent receiver operating characteristic curve(ROC)analysis.These results suggest that combining the mi RNA panel with pathological characteristics may provide a more accurate prognosis for HNSCC.mi RNA-548 k was highly expressed in the tumor tissues of patients with oral squamous cell carcinoma,which was significantly different from the adjacent tissues.In vivo and in vitro experiments,mi RNA-548 k mimics promoted tumor proliferation and invasion.The mi RNA-548 k inhibitor inhibited the proliferation and invasion of tumor cells.Bioinformatics analysis revealed that Lin-7C may be one of the downstream target genes of mi RNA-548 k.Further experimental results showed that the expression of mi RNA-548 k and Lin-7C in OSCC tumor tissues and cell lines was negatively correlated,and the direct interaction between the two was confirmed by fluorescence reporter gene experiment.The results of rescue and recovery experiments confirmed that the part of mi RNA-548 k could promote tumor proliferation and invasion by regulating Lin-7C.The PDX model showed that the mi R-548 k analogue could promote the growth of tumor tissue in vivo.Conclusion:(1)Our study demonstrates that the 7 mi RNAs panel reported here may be useful in making different prognoses and may improve the clinical management of patients with HNSCC.(2)These results indicated that mi R-548 k functions as an oncogene by directly targeting Lin-7C in OSCC.(3)mi R-548k/Lin-7C may be a potential therapeutic target and a novel prognostic biomarker for OSCC in the future.