| Objective : Laryngeal carcinoma is one of the most common head and neck malignancies,of which more than 95% are laryngeal squamous cell carcinoma.The incidence rate and prevalence rate of global laryngeal squamous cell carcinoma have increased by 12% and 24% over the past 30 years.This requires us to explore the mechanism of the occurrence and development of laryngeal squamous cell carcinoma and find new early diagnostic markers and therapeutic targets.Long non-coding RNA TUG1,which is the taurine up-regulated 1,has been found to be dysregulated in esophageal squamous cell carcinoma,osteosarcoma,breast cancer,liver cancer,colon cancer and non-small cell lung cancer,and plays an important regulatory role in the development of many malignant tumors.At present,the research on the expression and function of tug1 in laryngeal squamous cell carcinoma is not comprehensive,and the research on its mechanism is less.mi R-197-3p is located on human chromosome 1p13.3 and plays a role in promoting or inhibiting cancer development in different tumors.The expression and function of mi R-197-3p in laryngeal squamous cell carcinoma are not clear.The prediction of network information software suggests that mi R-197-3p may be the downstream target gene of TUG1 in laryngeal squamous cell carcinoma.This experiment is divided into three parts to detect the expression and function of TUG1 in laryngeal squamous cell carcinoma,the relationship between TUG1 and mi R-197-3p,the expression and function of mi R-197-3p in laryngeal squamous cell carcinoma,and the mechanism of TUG1/mi R-197-3p/CDK8 in laryngeal squamous cell carcinoma.Methods : 71 laryngeal squamous cell carcinoma tissues and 26 adjacent normal mucosa were selected.The expressions of TUG1,mi R-197-3p and CDK8 were detected by q RT-PCR.Combined with clinical data,the relationship between age,gender,lesion location,T stage,tumor differentiation,lymph node metastasis,TNM stage and the expression levels of TUG1 and mi R-197-3p in patients with laryngeal squamous cell carcinoma was analyzed.Kalan Meier analyzed the survival of patients with laryngeal squamous cell carcinoma with different expression levels of TUG1 and mi R-197-3p.Cox regression model was used to analyze the relationship between the expression of mi R-197-3p and the overall survival rate of patients with laryngeal squamous cell carcinoma.The cell function of laryngeal squamous cell carcinoma TU212 cell line was tested.The effects of TUG1 knockdown and mi R-197-3p overexpression on the proliferation of laryngeal squamous cell carcinoma cells were detected by CCK8 method.The effects of TUG1 interference and mi R-197-3p overexpression on cell cycle and apoptosis of laryngeal squamous cell carcinoma were detected by flow cytometry.Transwell was used to detect the effects of TUG1 knockdown and mi R-197-3p overexpression on the migration and invasion of laryngeal squamous cell carcinoma cells.The possible downstream target genes of TUG1 and mi R-197-3p in laryngeal squamous cell carcinoma were predicted by using network information software such as Starbase and targetscan.Pearson test was used to analyze the correlation between TUG1,mi R-197-3p and CDK8.Double luciferase reporter gene experiment confirmed that TUG1 and mi R-197-3p could directly bind to downstream target genes in laryngeal squamous cell carcinoma cells.The effects of TUG1/mi R-197-3p/CDK8 axis on cell cycle and proliferation of laryngeal squamous cell carcinoma were detected by flow cytometry cell cycle and CCK8 cell proliferation rescue experiment.Results:1.The expression level of TUG1 in laryngeal squamous cell carcinoma was higher than that in adjacent normal mucosa;The expression level of TUG1 is relatively higher in patients with low differentiation,high T-grade and TNM stage of laryngeal squamous cell carcinoma.The prognosis of patients with high expression of tug1 in laryngeal squamous cell carcinoma is relatively poor.TUG1 interference can inhibit the proliferation of laryngeal squamous cell carcinoma cells,induce apoptosis,make laryngeal squamous cell carcinoma cells appear S-phase block,and inhibit the migration and invasion of laryngeal squamous cell carcinoma cells.2.mi R-197-3p is low expressed in laryngeal squamous cell carcinoma,and its expression level is relatively low in patients with higher t grade and TNM stage and lymph node metastasis.The prognosis of patients with low expression of mi R-197-3p in laryngeal squamous cell carcinoma is relatively poor,which is an independent risk factor affecting the survival rate of patients with laryngeal squamous cell carcinoma.Overexpression of mi R-197-3p can inhibit the proliferation of laryngeal squamous cell carcinoma cells,induce apoptosis,block the cell cycle of laryngeal squamous cell carcinoma cells,and reduce the migration and invasion of laryngeal squamous cell carcinoma cells.3.Double luciferase reporter gene experiment confirmed that mi R-197-3p and TUG1,CDK8 and mi R-197-3p,could directly bind in laryngeal squamous cell carcinoma.CDK8 was overexpressed in laryngeal squamous cell carcinoma.There was a negative correlation between mi R-197-3p and CDK8 gene expression in laryngeal squamous cell carcinoma.mi R-197-3p inhibition could counteract the effect of TUG1 knockdown on the proliferation of laryngeal squamous cell carcinoma cells.Conclusion:1.TUG1 is overexpressed in laryngeal squamous cell carcinoma.TUG1 interference can inhibit the proliferation,migration and invasion of laryngeal squamous cell carcinoma cells,promote apoptosis and affect cell cycle.2.mi R-197-3p is the target gene of TUG1 in laryngeal squamous cell carcinoma;mi R-197-3p was low expressed in laryngeal squamous cell carcinoma;Overexpression of mi R-197-3p can inhibit the proliferation,migration and invasion of laryngeal squamous cell carcinoma cells.3.CDK8 is the target gene of mi R-197-3p in laryngeal squamous cell carcinoma;TUG1 regulates the proliferation of laryngeal squamous cell carcinoma cells through mi R-197-3p/CDK8. |
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