The Molecular Mechanism Of E IF5A-1 Gene In Cervical Cancer Induced By E6/HPV 16+ Gene

Objective:The vast majority of invasive cervical cancer associated with HPV infection,E6/HPV 16+gene plays an important role in cervical cancer cell activity,migration and proliferation,in this study,to investigate whether eIF5A-1 gene mediates a series of biological reactions such as proliferation and migration of cervical cancer cells induced by E6/HPV 16+gene,to explore the molecular mechanism of the occurrence and development of cervical cancer and to find potential new targets and theoretical basis for the clinical treatment of cervical cancer.Methods:Forty cases of cervical cancer tissues and adjacent normal cervical tissues were collected,and the expression levels of E6/HPV 16+protein and eIF5A-1 protein were detected by Western blot.Recombinant plasmids E6/HPV 16+plasmids,E6/HPV16+siRNA plasmids,recombinant plasmids eIF5A-1 and eIF5A-1 siRNA plasmids were transfected into human cervical cancer SiHa cell lines,or treated with the specific inhibitor cyclopyrrole(CPX).Real-time PCR and Western blot were used to detect the expression levels of E6/HPV 16+protein and eIF5A-1 gene.Immunohistochemistry was used to localize the expression of E6/HPV 16+and e IF5A-1.The apoptosis rate of cervical cancer cells was measured by flow cytometry.Methyl-thiazolidazole tetrazolium(MTT)colorimetry was used to detect the activity of cervical cancer cells.Transwell culture plate can convert the migration capacity of cervical cancer cells;The proliferation capacity of cervical cancer cells was detected by ~3H-thymine nucleotide involvement(~3H-TdR).Results:The expression level of E6/HPV 16+protein in cervical tissue was detected,and the results showed that the expression level of E6/HPV 16+protein was significantly up-regulated in 34 cases(85.0%)of cervical cancer tissues,while the expression level of E6/HPV 16+protein was up-regulated in only 2 cases(5%)of paracancer tissues.Moreover,the expression level of E6/HPV 16+protein was mainly located in the cytoplasm.Silencing E6/HPV 16+gene significantly reduced SiHa cell proliferation and migration ability,which was statistically significant compared with Negative siRN A group(p<0.01).SiHa cell proliferation activity of overexpressed E6/HPV 16+gene was significantly increased,showing a statistical difference(p<0.01).Apoptosis of SiHa cells was significantly increased in the transfected E6/HPV 16+siRNA plasmid(p<0.01).Compared with the empty group,the apoptotic number of SiHa cells transfected with E6/HPV 16+vector was significantly reduced,and the difference was statistically significant(p<0.01).Real-time PCR,western blotting and immunohistochemistry were used to detect the expression of eIF5A-1 protein in cervical tissues.The results showed that eIF5A-1 gene was highly expressed in cervical squamous cell carcinoma tissues,and eIF5A-1 protein was mainly located in the cytoplasm.Real-time PCR and western blot results showed that E6/HPV 16+gene could induce the expression of eIF5A-1 gene.The results of activity,migration and proliferation of cervical cancer cells showed that,compared with the Negative siRNA group,the activity,migration and proliferation of SiHa cells were significantly decreased after transfection with e IF5A-1 siRNA plasmid(p<0.05).Compared with the empty plasmid group,the activity,migration and proliferation capacity of SiHa cells were significantly increased after transfection with eIF5A-1 vector plasmid,showing a statistically significant difference(p<0.01).By preventing the maturation of eIF5A-1,the CPX drug reduced the activity,migration and cell proliferation of cervical cancer cells,and the difference was statistically significant(p<0.05).Cells that were silenced by eIF5A-1 gene or CPX-treated were 3.13 and 3.05 times more likely to induce apoptosis than those treated by E6/HPV 16+alone(p>0.01).Simultaneous detection of cervical cancer showed that there were many vacuoles in the cytoplasm,the cell nucleus was marginalized,part of the cell nucleus chromatin was aggregated,increased and presented marginalization,and apoptotic bodies were formed in the cell.Moreover,the activity,migration and proliferation of cervical cancer cells were significantly decreased,and the difference was statistically significant(p<0.01).Conclusion:E6/HPV 16+gene plays an important role in the tumorigenic transformation of cervical cancer cells.eIF5A-1 gene silencing or blocking hypusine enzyme activity effectively inhibits the viability,migration and proliferation of cervical cancer cells induced by E6/HPV 16+.Therefore,eIF5A-1 gene plays a key role in the cervical cancer cells induced by E6/HPV 16+and can become a potential intervention target for cervical cancer treatment.