Effect Of MicroRNA-101 On Human Laryngeal Squamous Cell Carcinoma By Targeting EZH2 And Its Clinical Significance

ObjectiveLaryngeal squamous cell carcinoma(Laryngeal squamous cell carcinoma,LSCC)is a common malignant tumor in the otolaryngology field.It is the second most common head and neck cancer in the body.The incidence rate is increasing,and the recurrence rate is high and the survival rate is low.New methods for treating LSCC have been studied at the genetic level.Micro RNA-101(mi R-101)is reported to be a tumor suppressor in a variety of malignancies including laryngeal squamous cell carcinoma(LSCC).However,the role and molecular mechanism of mi R-101 in the development of LSCC has not been fully elucidated.In order to further explore the new treatment plan of LSCC and improve the clinical survival rate,this study used LSCC cell line as the research object,and further explored the effect and mechanism of mi R-101 on LSCC through various experimental methods,as well as significant clinical significance.MethodsThe expression of mi R-101 and zeste homolog 2(EZH2)m RNA was detected by RT-q PCR.Cell viability was determined by MTS assay.Cell apoptosis was assessed by flow cytometry and commercial caspase-3 kit by detecting apoptosis index and caspase-3 activity.Western Blot assay was performed to examine the expression of EZH2 protein.Bioinformatics analysis was performed to predict the target genes that mi R-101,and the relationship between mi R-101 and target gene was further determined by luciferase assay and RNA immunoprecipitation(RIP)..Resultsmi R-101 expression was significantly down-regulated in the LSCC cell line.MTS assays showed that overexpression of mi R-101 inhibited proliferation of Hep-2 and TU-212 cells.Flow cytometry analysis showed that mi R-101 overexpression resulted in a significant increase in the apoptotic index in Hep-2 and TU-212 cells.Moreover,mi R-101 overexpression stimulated the activity of caspase-3,which further indicated the promotory effect of mi R-101 on cell apoptosis.Target Scan’s online site predicted the potential binding sites of mi R-101 in EZH2 3’UTR.Following luciferase assay,RNA immunoprecipitation(RIP)and Western Blot analysis disclosed that mi R-101 inhibits EZH2 expression through direct interaction.Rescue experiments showed that up-regulation of EZH2 attenuated mi R-101-mediated antitumor activity in LSCC cells.Conclusion1.Ectopic expression of mi R-101 inhibits cell proliferation and promotes apoptosis in LSCC.2.mi R-101 inhibits proliferation and stimulates apoptosis by targeting EZH2 in LSCC,laying a theoretical foundation for understanding the pathogenesis of LSCC and developing a novel biomarker for LSCC treatment.