Methylation Of The Whole Group Of Early Sporadic Colorectal Cancerspectral Feature Analysis And Screening Of Early Diagnosis Markers

Part OneEarly sporadic colorectal cancer genome Methylation spectrum characteristic analysisOBJECTIVE:Analysis of the genome-wide methylation spectra of DNA in Early sporadic colorectal cancer tumor tissue DNA,tumor patients stool extraction DNA and Normal rectal mucosa DNA.METHODS:Using whole genome high-throughput chip technology(Infmium 450KMethylation BeadChip methylation chip),The whole genome methylation spectrum of DNA was performed in Early sporadic colorectal cancer tumor tissue DNA,tumor patients stool extraction DNA and Normal rectal mucosa DNA.The data analysis and selection of the microarray data analysis software were carried out by the microarray analysis software(Significance Analysis of Microarrays,SAM).Attempt to construct the whole genome methylation spectrum information base of DNA in Early sporadic colorectal cancer tumor tissue DNA,tumor patients stool extraction DNA and Normal rectal mucosa DNA.And try to describe and compare their features.RESULTS:Original data of genome-wide methylation chip was analysised by microarray significant analysis(SAM).And screening,combined with biological analysis software(GO,KEGG)gene cluster analysis and gene biological pathways were analyzed,and finally the discovery of early sporadic knot colorectal tumour tissue DNA and tumor patients with fecal extract the genome-wide methylation profiles of DNA methylation distribution characteristics are similar,they and normal rectal mucosa genome-wide methylation spectrum of DNA methylation of distribution,there was a significant difference.Application of GO biological analysis software enrichment analysis to find 179 gene set in Early sporadic colorectal cancer tumor tissue DNA,tumor patients stool extraction DNA and normal rectal mucosa tissue DNA.There are differences between of them.And through the KEGG biological pathway analysis found 19 of signaling pathway in the early stage of sporadic colorectal carcinoma DNA,tumor patients stool extraction DNA and normal rectal mucosa tissue DNA,there were significant differences between the these fabric DNA.CONCLUSION:DNA methylation of DNA in early sporadic colorectal cancer tissues was similar to that of tumor patients stool extraction DNA.There were differences between the normal rectal mucosa tissues DNA with them in DNA methylation status of DNA methylation spectra.These abnormal DNA methylation sites may play an important role in the development and development of early sporadic colorectal cancer.They maybey imply that DNA methylation is a frequent event for early sporadic colorectal cancer.And It also shows that the development of early sporadic colorectal cancer is the result of the combined effect of multiple oncogenes or tumor suppressor genes.Part TwoScreening methylation microarraybased on methylation microarrayOBJECTIVE:By early sporadic colorectal cancer tissues DNA and feces of patients with tumor extract the raw data of genome-wide methylation profiles of DNA and normal rectal mucosa DNA methylation chip analysis,combined with multiple bioinformatics software screening,screening possible with gene expression and regulation of DNA methylation gene.METHODS:The whole gene methylation spectra of DNA were compared and analyzed in early sporadic colorectal cancer tumor tissue DNA,tumor patients stool extraction DNA and Normal rectal mucosa DNA.While the methylation gene was screened by the analysis of the software(SAM).At last these gene clusters and biological signal pathways of the gene were analyzed by using the differentially expressed genes of the gene into the biological information,and the DNA methylation groups used for mining functional properties.RESULTS:The analysis by bioinformatics software gene cluster,biological signal pathway on early stage of sporadic colorectal cancer tissues,feces DNA whole DNA gene methylation profiles,DNA methylation group express multiple differences,and normal tissue expression have 179 significant differences in DNA methylation.The application of KEGG signal pathway analysis,found that the expression of 19 differentially methylated genes involved in related signal pathway of multiple tumor.CONCLUSION:In the development of sporadic colorectal cancer,the possible number of tumor associated signaling pathways is affected by the abnormal methylation of DNA.These abnormal DNA methylation genes may be involved in the development and development of sporadic colorectal cancer.These aberrant DNA methylation gene also reflects early sporadic occurrence and development of colorectal cancer is a multiple signal pathways are involved in complex diseases,also reveals the early sporadic colorectal cancer is a multi gene involved in disease.This conclusion provides a reference for the molecular mechanism of early sporadic colorectal cancer.Part ThreeMethylation difference of differential gene and its diagnostic value for early sporadic colorectal cancer in tumor,feces and normal tissuesOBJECTIVE:To detect and analyze the methylation status of RP5 in the DNA of sporadic colorectal cancer tissues,tumor patients stool extraction DNA and Normal rectal mucosa DNA.Evaluation of the value of the application of gene RP5 in the screening and diagnosis of early sporadic colorectal cancer.METHOD:The tumor tissues of early sporadic colon cancer were collected in 30 cases,the stool specimen of colorectal cancer patients and 30 cases of normal rectal mucosa tissues were collected.Application of nested methylation specific PCR(NMSP)binding to agarose gel electrophoresis was used to test the difference expression of gene Rp5 in tumor tissue methylation,Using nested methylation specific PCR(NMSP)binds to the agarose gelelectrophoresis method to examine the differences in expression of genes Rp5 in sporadic node direct tumor tissue DNA and feces of patients with tumor extract DNA and normal rectal mucosa DNA methylation,and compare the difference between above of them.At last the sequencing of gene was sequenced by BSP sequencing.This study uses the software SPSS17.0 to do statistical analysis.If the measurement data,then the t test;if the count data,then uses ?2 analysis.RESULTS:There were 22 cases with methylation(73.3%)samples in tumor group of 30 cases,and there were 19 cases with methylation(63.3%)samples in faeces group of 30 cases,while there were only 2 cases with methylation(6.6%)samples in the control group of 30 cases by electrophoresis method.By using X2statistical analysis of the comparison between the data,the difference of expression of gene RP5 in the tumor group of methylation frequency(73.3%)was significantly higher than the control group methylation frequency(6.6%),there were significant differences between them(?2=29.584 P<0.001),while there was not differences between tumor groups and feces groups which their comparison of the methylation frequency of the expression.CONCLUSION:The methylation frequency of in the early sporadic colorectal cancer tumor tissue DNA and the methylation frequency of DNA in the stool extracted from the feces were significantly higher than that the methylation frequency of normal rectal mucosa comes fromm gene RP5.Differential gene RP5 may be a molecular marker for early screening and diagnosis of early sporadic colorectal cancer..Especially the detection of fecal DNA is likely to become a noninvasive detection method for early screening of fecal DNA and early diagnosis of sporadic colorectal cancer.