Effect Of LncRNA MAGI2-AS3 Regulating MiR-374a-5p/PTEN On The Biological Behavior Of Oral Cancer Cells

ObjectiveTo investigate the effect and mechanism of lncRNA MAGI2-AS3 in regulating miR-374a-5p / PTEN on the proliferation,cloning,cycle,invasion,migration and apoptosis of oral squamous cell carcinoma.Methods1.The qRT-PCR method was used to determine the expression of MAGI2 antisense RNA 3(MAGI2-AS3)in oral squamouscell carcinoma tissues and adjacent tissues,and to detect the expression of MAGI2-AS3 in oral squamous-cell carcinoma cells CAL-27,SCC15,HSQ-89 and normal oral mucosal cells h NOK;Transfection of MAGI2-AS3 overexpression vector in cancer cells,and transfection of negative control vector at the same time,MTT was used to determine cell proliferation,plate cloning test was used to detect cell clone formation ability,PI single staining method was used to detect cell cycle,Annexin V-FITC / PI double staining method detected apoptosis,and Transwell cells were used to detect cell invasion and migration.2.Bioinformatics software predicts the target gene of MAGI2-AS3,the targeting relationship between MAGI2-AS3 and miR-374a-5p was identified by the luciferase reporter system,and qRT-PCR method was used to determine oral squamouscell carcinoma cells transfected with MAGI2-AS3 overexpression vector miR-374a-5p expression level.3.qRT-PCR method was used to determine the expression of miR-374a-5p inoral squamouscell carcinoma tissues and adjacent tissues,and to detect miR-374a-5p expression in oral squamouscell carcinoma cells CAL-27,SCC15,HSQ-89 and normal oral mucosal cells h NOK;transfection of miR-374a-5p mimics and MAGI2-AS3 overexpression vectors inoral squamouscell carcinoma cells,transfection with mimics control,MTT to determine cell proliferation,plate cloning assay to detect cell clone formation ability,PI single staining method to detect cell cycle,Annexin V-FITC / PI double staining method was used to detect apoptosis,and Transwell chamber was used to detect cell invasion and migration.4.Bioinformatics software predicts miR-374a-5p target genes,the luciferase reporter system identifies the targeting relationship between miR-374a-5p and phosphatase and tensin homolog deleted on chromosome 10(PTEN),qRT-PCR and Western blot methods were used to determine the PTEN expression levels in oral squamouscell carcinoma cells transfection of miR-374a-5p mimics.5.The qRT-PCR method was used to determine the expression of PTEN in oral squamouscell carcinoma tissues and adjacent tissues,and to detect the expression of PTEN in oral squamouscell carcinoma cells CAL-27,SCC15,HSQ-89 and normal oral mucosal cells h NOK;Stained with miR-374a-5p mimics and PTEN overexpression vector,and transfected negative control vector at the same time.MTT was used to determine cell proliferation.Plate cloning experiment was used to detect cell clone formation ability.PI single staining method was used to detect cell cycle.Annexin V-FITC / PI double staining method was used to detect apoptosis,and Transwell chamber was used to detect cell invasion and migration.Results1.The expression level of MAGI2-AS3 in oral squamouscell carcinoma tissues was lower than that in adjacent tissues.The expression level ofMAGI2-AS3 in oral squamouscell carcinoma cells CAL-27,SCC15,and HSQ-89 was lower than that of normal oral mucosal cells h NOK;Comparing the cells,the transfection ability of the MAGI2-AS3 overexpression vector reduced the cell proliferation,cloning,invasion and migration ability,the cell G0 / G1 phase ratio increased,and apoptosis increased.2.MAGI2-AS3 targeted inhibition of miR-374a-5p expression in oral squamou-scell carcinoma cells.3.The expression level of miR-374a-5p in oral squamouscell carcinoma tissues was higher than that in adjacent tissues.The expression of miR-374a-5p and MAGI2-AS3 was negatively correlated in oral squamouscell carcinoma tissues,andthe expression level of miR-374a-5p in oral squamouscell carcinoma cells CAL-27,SCC15,HSQ-89 were higher than that of normal oral mucosal cells h NOK;compared with cells transfected with mimics control and MAGI2-AS3 overexpression vector,cells transfected with miR-374a-5p mimics and MAGI2-AS3 overexpression vector cell proliferation,cloning,invasion and migration increased,cell G0/G1 phase ratio decreased,and apoptosis decreased.4.miR-374a-5p targeted inhibition of PTEN expression in oral squamouscell carcinoma cells.5.The expression level of PTEN in oral squamouscell carcinoma tissues was lower than that in adjacent tissues.There was a positive correlation between PTEN and MAGI2-AS3 expression in oral squamouscell carcinoma tissues,and a negative correlation between PTEN and miR-374a-5p expression in oral squamouscell carcinoma tissues.The expression levels of PTEN in oral squamouscell carcinoma cells CAL-27 SCC15 and HSQ-89 were lower than those of normal oral mucosal cells h NOK;compared with cells transfected with miR-374a-5p mimics and negative control vectors,cells transfected with miR-374a-5p mimics and PTEN overexpression vectors proliferation,cloning,invasion,and migration were reduced,the G0/G1 phase ratio of cells increased,and apoptosis increased.ConclusionslncRNA MAGI2-AS3 regulates miR-374a-5p/PTEN to inhibit oral squamouscell carcinoma cell proliferation,cloning,invasion,migration,block the cell cycle,and induce apoptosis.