Circular RNA Hsa＿circ＿0015211 Promotes The Progression Of Laryngeal Squamous Cell Carcinoma And Its Impact On Patient Prognosis

Objective:Circular RNA(circular RNA,circRNA)is a type of single-stranded closed circular RNA linked end-to-end by covalent bonds.With the progress of research in recent years,more and more evidences have shown that the abnormal expression of circular RNA plays a crucial role in many diseases including cancer,but few reports were found about head and neck tumors.This study explored the expression of hsa＿circ＿0015211 in laryngeal squamous cell carcinoma(LSCC)tissue and its effect on clinicopathological indicators and prognosis of LSCC patients,and verified the promotion of hsa＿circ＿0015211 on human LSCC cell proliferation,migration and invasion effect.Methods:1.The differentially expressed hsa＿circ＿0015211 in human LSCC tissues and corresponding paracancerous tissues was screened out through the GEO database.2.Fluorescence in situ hybridization(FISH)was used to detect the expression of hsa＿circ＿0015211 in 67 human LSCC tissues and corresponding paracancerous tissues,and the relationship between its expression and clinicopathological indicators and prognosis of LSCC patients was analyzed.3.Use siRNA to knock down the expression of hsa＿circ＿0015211 in human LSCC cells,and verify the effect of hsa＿circ＿0015211 on its proliferation,migration and invasion by CCK8 experiment,clone formation assay,scratch healing assay and Transwell assay.4.The statistical software SPSS26.0 was used to conduct statistical analysis on the data,and P<0.05 was regarded as a statistically significant difference.Results:1.The expression of hsa＿circ＿0015211 was significantly increased in LSCC tissues compared with paracancerous tissues.2.The high expression of Hsa＿circ＿0015211 was correlated with the age of LSCC patients(x~2=3.963,P=0.046),pathological stage(x~2=6.041,P=0.014),pathological differentiation grade(x~2=8.323,P=0.004)and lymph node metastasis(x~2=4.219,P=0.040)were all positively correlated.Kaplan-Meier analysis showed that differential expression of hsa＿circ＿0015211affected the prognosis of LSCC patients.Compared with patients with low expression of hsa＿circ＿0015211,patients with high expression of hsa＿circ＿0015211 had a lower overall survival rate(P<0.05).Cox regression analysis showed that the expression of hsa＿circ＿0015211,clinicopathological stage and clinicopathological differentiation grade were independent risk factors affecting the prognosis of LSCC patients.3.The results of qRT-PCR experiments showed that in human LSCC cell lines AMC-HN-8 and TU177,the relative expression level of hsa＿circ＿0015211 in AMC-HN-8 was relatively high,so this cell was selected for subsequent knockdown experiments.4.The results of RT-PCR experiments showed that circular transcripts can only be amplified in the c DNA of AMC-HN-8 cells,and linear transcripts can be amplified in both c DNA and g DNA of AMC-HN-8 cells,verified the existence of hsa＿circ＿0015211 in human LSCC cells.5.After knocking down hsa＿circ＿0015211 in AMC-HN-8 cells,the results of q RT-PCR experiments showed that the expression level of hsa＿circ＿0015211 in the si-hsa＿circ＿0015211 group was significantly lower than that in the si-NC group,and the difference was statistically significant(P<0.05).6.The results of CCK8 experiment and plate colony formation experi-ment showed that the cell proliferation ability of the hsa＿circ＿0015211knockdown group was significantly lower than that of the control group,and the difference was statistically significant(P<0.05).7.The results of cell scratch healing assay and Transwell assay showed that the cell migration and invasion abilities of the hsa＿circ＿0015211knockdown group were significantly lower than those of the control group,and the difference was statistically significant(P<0.05).Conclusions:1.Hsa＿circ＿0015211 is highly expressed in LSCC tissues,which is correlated with age,disease pathological stage,pathological differentiation grade and lymph node metastasis were positively correlated.Patients with higher Hsa＿circ＿0015211 expression,higher pathological stage,and poorer pathological differentiation grade had poorer overall survival.2.Hsa＿circ＿0015211 can promote the proliferation,migration and invasion of human LSCC cells.It suggested that hsa＿circ＿0015211 might affect the progression of LSCC.