The Influence Of Hsa_circ₀112879 On The Biological Characteristics In Oral Carcinoma Cell Lines

Purpose:Oral squamous cell carcinoma is a common malignancy in the world with extremely high mortality and recurrence rates.At present,the mechanism of occurrence and development of oral squamous cell carcinoma has not been studied clearly.Circular RNAs?CircRNAs?are a class of non-coding RNAs that contain continuous stable loop structure without 5?-3?ends and poly A tail.Studies have found that many circRNAs play an important role in the development of cancer and may serve as novel biomarkers for clinical diagnosis.The purpose of this study was to detect the expression of hsa_circ₀112879 in oral squamous cell carcinoma cell lines and to explore its role in the development of oral squamous cell carcinoma and related molecular mechanisms.Methods:The expression of hsa_circ₀112879 in five cell lines was detected by qRT-PCR.The appropriate cell lines were selected for low expression experiments.The siRNAs were transfected into selected cells and the effect of siRNA interference was detected by qRT-PCR.Proliferation assay was used to detect the changes of cell proliferation.Cell migration and invasion were evaluated by scratch and Transwell assay.Cell apoptosis rate was determined by Hoechst staining and Annexin V-FITC/PI flow cytometry.The expression of apoptotic proteins BAX and BCL-2 in cells was detected by Western blot.Results:The expression of hsa_circ₀112879 in five cell lines?HOK,SCC9,SCC15,SCC25,CAL27?were detected by qRT-PCR.The results showed that the expression levels of SCC15 and CAL27 were the lowest.The results of qRT-PCR showed that the expression level of circRNA₀112879 in oral squamous cell carcinoma cell lines SCC15 was decreased by about 85%compared with the control group,while the expression level of hsa_circ₀112879 in oral squamous cell carcinoma cell lines CAL27was lower than that of the control group75%(t_SCC15=81.38,P_SCC15=0.0002,***P<0.001;t_CAL27=27.61,P_CAL27=0.0013,**P<0.01).The results of CCK-8 proliferation assay showed that the proliferation of SCC15 and CAL27cells gradually increased as compared with the control group as the cell growth time went by?*P<0.05?.The results of EdU proliferation assay showed that the proliferation of SCC15 and CAL27cell lines was higher than that of control group(t_SCC15=5.18,P_SCC15=0.031,t_CAL27=5.68,P_CAL27=0.03,*P<0.05).Scratch results showed that:the interference group than the control group,the number of scratched cells increased and the migration distance increased.The results of Transwell migration showed that the numbers of cells passing through the fundus in SCC15 and CAL27 cells were significantly increased compared with the control group(t _SCC15=12.00,P_SCC15=0.0069,**P<0.01;t_CAL27=11.67,P_CAL27=0.0073,**P<0.01).The results of Transwell invasion assay showed that the numbers of cells passing through the chambers were significantly increased in the SCC15 and CAL27 cells compared with the control group(t_SCC15=12.85,P_SCC15=0.006,**P<0.01;t_CAL27=9.70,P_CAL27=0.011,*P<0.05).The results of Hoechst staining showed that the number of apoptotic cells in SCC15 and CAL27 cells was significantly decreased compared with the control group(t_SCC15=12.85,P_SCC15=0.006,**P<0.01;t_CAL27=9.70,P_CAL27=0.011,*P<0.05).Annexin V-FITC/PI dual-label flow cytometry results showed that the apoptosis rates in the interference group and the control group in SCC15 and CAL27 cells were 0.97%,1.48%,0.45%,and 1.54%,respectively.The results were statistically significant?*P<0.05?.The results showed that low expression of hsa_circ₀112879 inhibited the apoptosis of SCC15 and CAL27 cells.Western blot showed that the expression of BAX in SCC15 and CAL27cells was significantly decreased and the expression of BCL-2 was significantly increased.Conclusion:By interfering with the expression level of hsa_circ₀112879,the proliferation,migration,invasion and apoptosis of SCC15 and CAL27 in oral squamous cell carcinoma can be enhanced.Hsa_circ₀112879 may play a role in inhibiting the develpoment of oral squamous cell carcinoma.