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Application of target gene DNA methylation as molecular markers in preparation of kit for distinguishing colorectal tissue canceration progress

A technology of molecular markers and target genes, which is applied in the field of preparation of kits for distinguishing colorectal tissue cancer progression, can solve the problem of few researches on markers of colorectal tissue cancer progression, and achieve high specificity and high sensitivity Effect

Active Publication Date: 2019-09-27
ZHEJIANG UNIV
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Problems solved by technology

[0010] However, the above studies are all aimed at molecular markers for the diagnosis of colorectal cancer, and there are few studies on markers for distinguishing the progression of colorectal cancer; moreover, the occurrence and development of colorectal cancer is a multi-stage long process, Studies have found that nearly 95% of colorectal cancers are evolved from polyps, and it usually takes 5 to 10 years to go through the process of normal mucosa → hyperplasia → adenoma formation → adenoma canceration

Method used

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  • Application of target gene DNA methylation as molecular markers in preparation of kit for distinguishing colorectal tissue canceration progress
  • Application of target gene DNA methylation as molecular markers in preparation of kit for distinguishing colorectal tissue canceration progress
  • Application of target gene DNA methylation as molecular markers in preparation of kit for distinguishing colorectal tissue canceration progress

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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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Abstract

The invention discloses an application of target gene DNA methylation as molecular markers in preparation of a kit for distinguishing colorectal tissue canceration progress. The target gene DNA methylation is one or more of COL25A1, GALR1, INHBA and HKDC1 genes. The invention finds for the first time that the DNA methylation molecular markers shown in SEQ ID NO.1-4 can be used for distinguishing the colorectal tissue canceration progress, an obtained model has better identification and diagnosis efficiency, and AUC is 0.662-0.734. The AUC of the model is 0.764 after four target gene DNA methylation markers are combined. The DNA methylation molecular markers have the characteristics of high sensitivity and high specificity when distinguishing rectal polyp and colorectal cancer and can assist in monitoring the process of rectal polyp to colorectal cancer.

Description

technical field [0001] The invention relates to the technical field of clinical examination and diagnosis, in particular to the application of DNA methylation of a target gene as a molecular marker in the preparation of a kit for distinguishing colorectal tissue cancer progression. Background technique [0002] Colorectal cancer (CRC) is one of the most common malignant tumors of the digestive system. According to statistics from the International Agency for Research on Cancer (IARC), there were approximately 1.8 million new cases of colorectal cancer in the world in 2018, ranking second in women and third in men. In recent decades, with the rapid development of my country's social economy, "Westernization" has brought about changes in lifestyle and diet structure. The incidence of colorectal cancer in my country has an obvious upward trend and is characterized by younger age. Colorectal cancer has become a major public health problem that seriously threatens the health of ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886
CPCC12Q1/6886C12Q2600/154
Inventor 陈坤金明娟顾思萌林淑娟钱桑妮王建炳唐梦龄李其龙
Owner ZHEJIANG UNIV
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