The Function Of N~6-methyladenosine RNA Binding Protein 2(YTHDF2)in Cervical Cancer Cells

Background:Cervical cancer is a common cause of death in gynecological malignancies.Although the clinical efficacy has been improved through surgery,chemotherapy,radiation therapy and targeted therapy,the prognosis of patients with cervical cancer is still unsatisfactory,which seriously endangers women's life,therefore,it's the key to find a new effective therapeutic target.N~6-Methyladenosine(m~6A)is the most common epigenetic modification of RNA in eukaryotes.Dysregulation of its main related proteins(writers,erasers and readers)contributes to the development of cancer.Objective:To observe the expression difference of YTH domain N~6methyl adenine(m~6A)RNA binding protein 2(YTHDF2)in cervical cancer tissue and normal cervical tissue.To investigate the effects of YTHDF2knockdown on the proliferation and apoptosis of human cervical cancer cells,providing evidence in vitro on the development of cervical cancer.To establish the relationship between cervical cancer and m~6A methylation modification.Methods:Human Protein Atlas database was used to analyze the expression of YTHDF2 in cervical cancer and its relationship with the survival of cervical cancer patients.Immunohistochemistry was performed to analyze protein expression of YTHDF2 in 31 cervical cancer tissue samples and 31 normal cervical tissue samples.Then short hairpin RNA(shRNAs)targeting YTHDF2 were designed and cloned into lentivirus expression vector pLKO.1.Furthermore,HeLa and SiHa cervical cancer cells were transfected with pLKO.1-shRNA to knockdown the expression of YTHDF2,which was confirmed by the detection of mRNA and protein expression using Real-Time quantitative PCR and Western blotting.Furthermore,cell proliferation was observed by CCK-8 assay and colony formation assay.Also,cell apoptosis were examined by flow cytometry.Results:Immunohistochemistry analysis showed that the expression of YTHDF2 in cervical cancer tissues was higher than that in normal cervical tissues.The higher expression of YTHDF2 was correlated with the shorter survival time in cervical cancer according to The Human Protein Atlas database.Cervical cancer cells stably expressing YTHDF2-shRNA were successfully established.Furthermore,YTHDF2 knockdown significantly inhibited the cell proliferation and induced cell apoptosis in both HeLa cells and SiHa cells after infection with lentivirus carrying shRNA1 and shRNA2 of YTHDF2 compared with those infected with lentivirus of empty vector.Conclusion:YTHDF2 is highly expressed in cervical cancer tissues,and knockdown of YTHDF2 in cervical carcinoma HeLa calls and SiHa cells significantly inhibits cell proliferation and promotes apoptosis.