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===Genome-wide DNA methylation profile comparison between solid tissue and circulating free DNA methylation in plasma=== All the assumption of we can used DNA methylation in free-cell circulating DNA methylation is based on the methylation status of solid cancers can be released into plasma without any specific selection, thus, the aberrant DNA methylation fragment or biomarkers identified in solid tissues can be used to be the target for cancer non-invasive diagnosis, screening or prognosis surveillance. We then constructed 15 genome-wide DNA methylation profiles for the paired plasmas and solid tissues from same cancer patient with RRBS assay to check whether DNA methylation fragments derived from cancer cells were released into the blood randomly or selectively. 5 paired lung cancer, colon cancer and pancreatic cancer solid tissues and plasma were enrolled in this section and single-base methylome were established by RRBS as mentioned above. Correlation analysis based on 56046 MHL regions showed there is just weak correlation between genome-wide DNA methylation of solid tissues and plasmas. The average correlation between tissue and plasma was 0.50 (95%CI: 0.44-0.56)(Figure). Whatβs more, only about 28% (95%CI: 25%-31%) regions were simultaneously methylated both in tissues and plasma. Even considering different threshold to define the methylation status and transferring the continuous MHL to binary DNA methylation status, the maximum co-methylation ratio was still less than 63% (Figure). These evidence showed the dramatically significant heterogeneity between solid and plasma DNA methylation profile and provide the importance that we should validate all the biomarkers identified in solid tissues in cell-free circulating plasma DNA to guarantee the validity of the biomarkers. 25204 regions which was methylated in sample pairs for at least one time were selected to evaluate the selectivity of hyper-methylated DNA fragments from solid tissues to plasma. The results showed the process of the releasing were non-random, with at least 1590 fragment were significantly prefer-selected (P<1.98*10-6, binomial test). These fragments were located in the regions of 1190 genes. Function enrichment analysis showed these genes were significantly associated with cancer relevant biological functions (Table), including embryonic morphogenesis, regulation of transcription, neuron differentiation, regionalization, tissue morphogenesis ,transcription factor activity, sequence-specific DNA binding ,transcription regulator activity. [[File:1155.tm.png|400px]]
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