Application of target gene DNA methylation as a molecular marker in the preparation of a kit for discriminating colorectal cancer progression
A technology of molecular markers and target genes, which is applied in the field of preparation of kits for identifying colorectal cancer progression, can solve the problems of few researches on markers of colorectal cancer progression, and achieve good diagnostic efficiency and high specificity Sexual, high-sensitivity effects
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Embodiment 1
[0033] The screening process of methylation markers is divided into three stages, which are screening stage, verification stage I and verification stage II.
[0034] (1) In the screening stage, IlluminaInfinium Methylation EPICBeadChip (850K) was used to screen positive sites, and 12 cases of colorectal cancer (cancer and paracancerous tissue) and 3 cases of colorectal polyp (polyp and parapolypoid tissue) were sent for inspection. ) specimen. A total of 5,616 differentially methylated sites were screened out between cancer and adjacent normal tissues, and a total of 183 differentially methylated sites were found in cancer tissues / polyps.
[0035] Finally, at the tissue level, 13 CpG sites were screened for validation phase I analysis.
[0036] (2) During the verification phase I, 46 cases of colorectal cancer were detected by pyrosequencing method to detect the methylation level of cancer / adjacent normal tissues, and the experimental results were consistent with the microarr...
Embodiment 2
[0041] 1. Research object
[0042] Tissue samples included 142 cases of colorectal polyps (including 13 cases of hyperplastic polyps and 129 cases of adenoma) and 302 cases of colorectal cancer and paired normal tissue samples.
[0043] The cases of colorectal polyps were the cases of intestinal polyps in the colorectal cancer screening cohort in Jiashan County, and the cases of colorectal cancer were from the confirmed cases of anorectal surgery in Shaoxing People's Hospital. All subjects were pathologically confirmed and those with a history of preoperative radiotherapy and chemotherapy, a history of familial adenomatous polyps, and a history of colorectal cancer were excluded. The study was reviewed by the Ethics Committee of Zhejiang University School of Medicine and informed consent was obtained from the subjects.
[0044] 2. Specimen collection
[0045] All the included colorectal cancer cases were resected and collected their corresponding cancer tissues and adjacent ...
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