The Role And Molecular Mechanism Of DIAPH3 In Cervical Cancer Cell Migration And Invasion

BackgroundCervical cancer is one of the most common malignant tumors for women.At present,although radiotherapy,chemotherapy and surgery are widely used in the treatment of cervical cancer,distant metastasis is the most common cause of death of cervical cancer patients in the progression of cervical cancer.In recent years,it has been found that DIAPH3 is involved in the development of lung cancer,liver cancer,pancreatic cancer,esophageal cancer and gastric cancer.Linling Wan et al.found that DIAPH3 inhibits the proliferation of cervical cancer cells through the m TOR signaling pathway,but the role and molecular mechanism of DIAPH3 in cervical cancer migration and invasion have not been reported.The research group planned to use gene transfection and in vitro invasion and migration experiment to detect the role of DIAPH3 in the invasion and migration of cervical cancer cells,so as to provide a new molecular target for the diagnosis and treatment of cervical cancer.ObjectivesIn this study,four strains of human cervical cancer cells,namely Si Ha,Hela,Caski and C33 A,were selected to analyze the expression of DIAPH3,and to explore the effects of DIAPH3 on the migration and invasion of cervical cancer cells and their molecular mechanisms.Methods1.Gene expression profiling interactive analysis（GEPIA）was used to analyze the expressions of DIAPH3 m RNA in cervical cancer tissues online.2.The expression of DIAPH3 protein and m RNA in four cervical cancer cells of Si Ha,He La,Caski and C33 A and human cervical epithelial immortem cell line H8 were detected by protein blotting（Western blot）and real-time PCR（real-time quantitative PCR,RT-PCR）.3.Cervical cancer cells were transfected with DIAPH3 si RNA interference fragment and overexpressed plasmid,respectively,and transfection efficiency was verified by Western blot and q PCR.4.The effects of interference/overexpression of DIAPH3 on the migration and invasion of cervical cancer cells were examined by scratch healing assay and Transwell chamber invasion assay.5.The pathway related to cervical cancer and DIAPH3 was analyzed by Gene Set Enrichment Analysis（GSEA）.Western blot was used to detect the protein expression levels of MMP2,MMP9,p-ERK and p-AKT in cervical cancer cells after interference/overexpression of DIAPH3.Results1.GEPIA database analysis showed that the expression level of DIAPH3 m RNA in cervical cancer tissues was significantly higher than that in adjacent cervical cancer tissues（P<0.05）.2.Western blot and QRT-PCR results showed that the expression of DIAPH3 in four cervical cancer cells was markedly higher than that in human cervical epithelial immortem cell line H8（P<0.05）.3.After interfering with DIAPH3 in Si Ha and Hela cells,the expression of DIAPH3 protein and m RNA was significantly lower than that in the control group（P<0.05）;After overexpression of DIAPH3 in Caski and C33 A cells,the expression of DIAPH3 protein and m RNA were significantly higher than that those in the control group（P<0.05）.4.The results of scratch healing experiment showed that after interfering with DIAPH3 in Si Ha and Hela cells,the cell mobility was significantly decreased compared with the control group（P<0.05）;After overexpression of DIAPH3 in Caski and C33 A cells,the cell mobility was significantly increased compared with the control group（P<0.05）.5.Transwell chamber invasion experiment showed that after interfering with DIAPH3,the invasion numbers of Si Ha and He La cells were significantly decreased compared with the control group（P<0.05）;After overexpression of DIAPH3,the invasion number of Caski and C33 A cells was significantly increased compared with the control group（P<0.05）.6.GSEA analysis results showed that DIAPH3 was closely related to migration and invasion（P<0.05）.Western blot results showed that after interfering with DIAPH3,the protein expressions of MMP2,MMP9,p-ERK and p-AKT proteins in Si Ha and Hela cells were significantly lower than those in control group（P<0.05）;After overexpression of DIAPH3,the protein expression of MMP2,MMP9,p-ERK and p-AKT in Caski and C33 A cells were significantly higher than those in the control group（P<0.05）.ConclusionsDIAPH3 promotes the migration and invasion of cervical cancer cells,and its molecular mechanism is related to the up regulation of MMP2,MMP9,p-ERK and p-AKT protein expression.