Screeing Of Prostate Cancer Methylation Associated Genes And Research The Relationship Between Candidate Gene VWA1 And Prostate Cancer

Objectives:Our research was to unearth some candidate genes that related with prostate cancer methylation using the data-mining methods of bioinformatics, and further to explore the relationship between the methylation statuses of gene VWA1 with prostate cancer.Methods:We applied the data-mining method of bioinformatics to analyze the database of NCBI/GEO in order to find some candidate genes that related with the methylation of prostate cancer. And then we selected one candidate gene VWA1 as a target gene. We verified this gene using the prostate methylation datasets from TCGA database. We applied BSP in benign prostatic hyperplasia tissue (BPH) and prostate cancer blood to investigate their difference. The BPH blood and prostate cancer blood were brought into according to the pathological biopsy outcome. We extracted DNA from blood samples of BPH and prostate cancer, and then qRT-PCR experiments were conducted to test the expression of VWA1 in each sample. In qRT-PCR experiments ((2-△△Ct) mean values±standard errors) were difined to stand for the level of espression. We compared the methylation ratio and the expression levels in order to find a new marker that associated with prostate cancer in blood. Excel、R packssage and SPSS 16.0 for windows were used for analyzing the data.Result:1. Outcome of bioinformatics(1) After filtraing 7 datasets were brought into our research scope according to our inclusion and exclusion criteria.(2) After calculating using fold change method and adjusted by expression data, we found 7 genes CELSR2、DNTTIP2、IL12RB2、OMA1、OXCT2、 PHC2 and VWAl were related with benigh prostate cancer; MYCBP、OMA1、 PPT1、PRKACB、ST7L and TMEM39B were related with metastatic prostate cancer. All of the 13 genes had statistically significant values.(3) We screened the above 13 genes in a epigenetic database, and found 9 genes MYCBP、PPT1、PRKACB、ST7L、TMEM39B. CELSR2. IL12RB2, PHC2 and VWAl had not been researched. After methylation sites prediction we found, PPT1 had 1 potential methylation site, CELSR2 had 5 potential methylation sites, IL12RB2 had one potential methylation site and VWAl had 2 potential methylation sites.In conclusion, we found PPT1 was related with metastatic prostate cancer, CELSR2, IL12RB2 and VWA1 were related with normal position prostate cancer.2. Outcome of TCGA database The methylation level of VWA1 was statistically significant, P value was 1.65E-11, and to analyze the 374 PCa samples and 52 BPH samples, we came to a conclusion that the expression level was significant P value was 0.032.3. Experiment results(1) Outcome of BSPThe length of gene VWA1 is 503bp, containing 82 CG locis. All of the samples that we analyzed was 12. To compare the two outcomes we could see, the proportion of methylation of VWA1 in prostate cancer samples was higher, and in benign prostate hyperplasia samples was lower. The difference of the methylation status between the two groups was statistically different (P=0.002).(2) Outcome of qRT-PCRWe used the relative expression ((2-△△Ct)mean value±standard error) to stand for the expression of gene VWA1 in different samples.In this part, expression of VWA1 was higher in benign prostate hyperplasia samples, and in prostate cancer samples it was lower. The difference of the relative expression between the two groups was statistically different (P=0.014).We can come to a conclusion from the two experiments, when the methylation status of VWA1 was higher the relative expression would be lower, and the methylation status of VWA1 lower its relative expression would be higher. The methylation status of VWA1 has a negative relation with its relative expression, the Pearson Correlation is -0.683. Thus, we could deduce that the methylation status of VWA1 might have a closely relationship with prostate cancer.Conclusion:PPT1 was found to be related with the metastatic prostate cancer, and CELSR2, IL 12RB2 and VWA1 were related with primary prostate cancer. Methylation status of VWA1 may be closely associated with prostate cancer.