Correlation Between DNA Methylation Of TP63 Gene Promoter And Cervical Cancer

Objectives To investigate the DNA methylation status of the TP63 gene promoter region in cervical cancer tissues,the m RNA expressions of TP63,survivin,and RUNX2,and the binding of RUNX2 to specific methylation sites in the TP63 gene promoter between two groups of cervical tissues.Correlation between DNA methylation status of TP63 gene promoter in cervical cancer tissues and clinicopathological parameters also was analysed.Methods A total of 85 cervical specimens were collected from the Tangshan Worker's Hospital from September 2015 to September 2018.The specimens were selected according to the inclusion and exclusion criteria of the specimens from patients with CC.Pathology of specimens was confirmed by an experimental staff and a hospital pathologist.The grades and types included 40 patients with cervical cancer and 45 patients with benign lesions undergoing hysterectomy.The status of DNA methylation and the m RNA expressions in CC and age-matched Control tissues(N=3 for each group)were screened by whole-genome DNA methylation analysis(WGBS)and RNA sequence.Bisulfite sequencing(BSP)and real-time quantitative PCR(q PCR)for verification on DNA methylation and m RNA expression of RUNX2,TP63,survivin in TP63 gene in cervical tissues.The binding of RUNX2 to the TP63 gene promoter in cervical tissues was detected by CHIP,and the relationship between DNA methylation status of TP63 gene and pathological parameters of SCC was also analyzed.Results 1 WGBS and RNA-seq screening revealed that elevated DNA methylation levels of TP63 promoter are associated with decreased m RNA expression.2 The BSP results confirmed that compared with Control,the DNA methylation rate of the-688 Cp G site in the promoter region of the TP63 gene in the CC group was significantly increased,and the difference was statistically significant(77.5%(31/40)vs.31.1%(14/45),P<0.05).3 q PCR results showed that compared with control,the expression of RUNX2 and survivin m RNA was up-regulated and the expression of TP63 m RNA was down-regulated in CC group(all P<0.05).4 Chromatin immunoprecipitation(Ch IP)confirmed the presence of transcription factor RUNX2 binding sequence in the promoter region of TP63 gene in CC group,and DNA methylation at-688 Cp G site affected the binding of RUNX2.5 Correlation analysis showed that DNA methylation of TP63 gene was associated with tumor size,pathological grade,and clinical stage(all P<0.05),regardless of age(P>0.05).Conclusions 1 DNA methylation of the TP63 gene promoter inhibiting TP63 expression is involved in the carcinogenesis and development of cervical cancer.2 TP63 is expected to be an important prognostic indicator for clinical evaluation of cervical cancer patients and a key molecular target for cervical cancer treatment.Figure6;Table5;Reference 148...