To Initially Explore The Role Of DNMT3a/3b And P16 Methylation In Cervical Cancer

Background:Cervical cancer(CC)is one of the common gynecological malignancies in women,and the pathogenesis is still unclear.The tumor suppressor gene p16 is a factor that has been widely studied.It has been found that methylation of Cp G island in the promoter region of the p16 gene can inactivate the p16 gene and cause abnormal cell proliferation and participate in the development of the tumor.DNA methyltransferases(DNMTs)affect gene expression by regulating the methylation level of the Cp G island in the gene promoter region and are key enzymes that catalyze the methylation reaction.DNMT3a/3b was found to be highly expressed in a variety of malignant tumors.By inducing methylation of Cp G island in the promoter region of anti-oncogenes,DNMTs inactivate anti-oncogenes and inhibit the expression of these genes in cells,which causes the imbalance of the cell cycle and promoting the proliferation and metastasis of cancer cells,leading to the development and progress of cancer.In thisstudy,the methylation status of the Cp G island of the P16 gene was measured in cervical cancer cells and the changes in the methylation status of the Cp G island of the P16 gene were measured by regulating the expression of DNMT3a/3b,to explore the roles of both and their possible regulatory mechanisms in the occurrence and development of cervical cancer,and to bring new ideas for precisely targeted drug therapy of cervical cancer in the future.Objective:To understand the methylation status of the promoter region of the Cp G island in the p16 gene in cervical cancer cell lines,and to determine the changes in the methylation status of the Cp G island of the P16 gene by silencing the expression of DNMT3a/ 3B in cervical cancer cells,to explore and analyze the role of methylation of the P16 gene in cervical cancer cells.Methods: 1.Two cell lines were cultured: 1)cervical cancer cell line Hela;2)normal cervical epithelial cell line ECT-1/E6E7;2.Methylation-specific PCR(MSP)was used to determine the methylation status of Cp G island in the p16 promoter region in both cell lines;3.After silencing the expression of DNMT3a/3b,changes in the methylation status of the Cp G island of the p16 gene were determined.Results:Cp G island Methylation at the promoter of the p16 gene was positive in Hela cells and negative in ECT1/E6E7 cells;2.p16 methylation was positive in the control group and the pc DNA3.1-NC transfected group,and p16 was partially methylated in the pc DNA3.1-DNMT3a/3b sh RNA transfected group.Conclusions:1.Cp G island Methylation at the promoter of the p16 gene was present in cervical cancer cells,while no methylation changes were observed in normal cervical cells.2.Silencing of DNMT3a/3b expression decreased the p16 methylation level,suggesting that DNMT3a/3b may affect the methylation level of the Cp G island in the p16 promoter,leading to cervical carcinogenesis.