Correlation Between Methylation Of ZNF772 Promoter Region And Cervial Cancer

Objectives To investigate the status of DNA methylation in the promoter region of ZNF772 gene and its m RNA and protein expression,function and clinical significance in cervical cancer tissues,and to verify the correlation between DNA methylation status in ZNF772 promoter and its expression.Methods From September 2015 to September 2018 in Tangshan Worker's hospital,a total specimens of 85 female patients including 40 cervical squamous-cell carcinoma(SCC group)and 45 benign diseases of uterus(control)were admitted to our hospital,the pathological report was.All cases were diagnosed by the chief pathologist of pathology department according to the inclusion and exclusion criteria.3 SCC tissues specimen and 3 age-matched normal cervical tissues were selected for screening hyper-methylation and lower transcripts using whole-genome bisulfite sequencing(WGBS)combined with RNA sequencing(RNA-seq);Real-time quantitative polymerase chain reaction(RT-q PCR)and bisulfite sequencing polymerase chain reaction(BSP)methods were performed to demonstrate screening results in 40 SCC tissues and 45 normal cervical tissues including DNA methylation status and m RNA expression of ZNF772;its protein expression was also detected by immunohistochemistry.The correlation between DNA methylation in ZNF772 promoter and its m RNA expression as well as clinicopathological parameters were analyzed.Results 1 There was no significant difference in the basic information(age,menarche age,menstrual cycle,pregnancy times,abortion times,smoking,drinking)between each group(P>0.05).2 The results of WGBS combined RNA-seq detection show that elevated DNA methylation level in the ZNF772 promoter is associated with down-regulated RNA expression(P>0.05);3 The results of RT-q PCR demonstrated that ZNF772 m RNA expression level in SCC was indeed decreased compared with the control,the difference was statistically significant(1.35 vs.0.43,t=8.351,P=0.016);4 BSP results also confirmed that compared with the control group,the DNA methylation rate of ZNF772 promoter(-420,-422)in SCC was significantly higher,the difference was significant(-420 site: ?2=8.566,P=0.038;-422 site: ?2=6.332,P=0.043);5 JASPAR website was used to predict the ZNF772 promoter sequence and transcription factor binding site,The results showed that the-420,-422 locus coincided with KLF5 and KLF4 cis acting elements,suggesting that the sites might affect the transcription level of ZNF772 associated with these two transcription factors;6 Immunohistochemical analysis further proved that ZNF772 expression localized in nuclei(the nucleus was brownish yellow granules)as a nuclear protein elicited a transcription function.Compared with the control group,the positive expression of ZNF772 in SCC decreased significantly(81.23% vs.10.18%,t=3.802,P=0.005);7 The spearman correlation analysis showed that in the control group,there was no difference in the m RNA expression level between methylation and unmethylation in-420 Cp G locus(1.18 vs.0.92,t=0.328,-420P=0.375);there was also no correlation between the m RNA expression and DNA methylation modification in-420 Cp G site of ZNF772(r-420=-0.089,P-420=0.263);In SCC,there was a significant difference of m RNA level between the methylation and unmethylation at-420 Cp G locus of ZNF772(0.31 vs.1.38,t=7.516,-420P=0.024);there was a negative correlation between the m RNA expression of ZNF772 and DNA methylation modification at-420 Cp G locus(r-420=-0.350,P-420=0.045);8 The correlation analysis of clinicopathological parameters in cervical cancer confirmed that: the DNA methylation of ZNF772 gene is related to tumor size,pathological grade and clinical stage(all P<0.05),and not related to patient's age(P>0.05).Conclusions 1 The increased DNA methylation rate of ZNF772 gene,and the decreased m RNA and protein expression human cervical cancer tissues.2 DNA hypermethylation in the promoter region of ZNF772 gene was related to the expression of m RNA and protein,involved in the occurrence and development of cervical cancer.3 Combined with clinical statistics,we found that DNA methylation of ZNF772 promoter region was related to tumor size,WHO pathological grade and FIGO clinical stage.Figure5;Table7;Reference131...