Application of DNA (deoxyribonucleic acid) binding site CTCF_13 of multifunctional transcriptional regulation factor CTCF

A technology of transcriptional regulatory factors and binding sites, applied in the application field of the DNA binding site CTCF_13 of the multifunctional transcriptional regulatory factor CTCF

Inactive Publication Date: 2017-09-12
FIRST AFFILIATED HOSPITAL OF KUNMING MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The DNA binding site of CTCF is ubiquitous in the human genome, there are about 50,000 to 60,000, and each site recognizes about 34 nucleotides; so far, t

Method used

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  • Application of DNA (deoxyribonucleic acid) binding site CTCF_13 of multifunctional transcriptional regulation factor CTCF
  • Application of DNA (deoxyribonucleic acid) binding site CTCF_13 of multifunctional transcriptional regulation factor CTCF

Examples

Experimental program
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Effect test

Embodiment 1

[0023] Example 1: Screening of the DNA Binding Site of the Multifunctional Transcription Regulator CTCF

[0024] 1. Sample collection

[0025] From April 2014 to August 2016, 187 cases of fresh tumor tissues and 84 cases of corresponding normal tissues (more than 6cm from the tumor edge) were collected from the First Affiliated Hospital of Kunming Medical University from April 2014 to August 2016; 108 cases of adenoma tissue. A total of 379 samples were taken, including 84 cases of normal tissues and 295 cases of tumor tissues; the clinical data of the above samples are shown in Table 1:

[0026] Table 1 Sample information table

[0027] .

[0028] 2. Genomic DNA extraction and bisulfite conversion

[0029] Genomic DNA was extracted using the QIAamp DNA Mini Kit (Product No. 51304) kit from QIAGEN, and all operations were performed according to the instructions of the kit. Bisulfite conversion was performed using the EpiTect Fast DNA Bisulfite Kit (Catalog No. 59826) fr...

Embodiment 2

[0056] Example 2: Enlarging the sample size for further screening of 10 loci

[0057] In order to improve the specificity and sensitivity of the screened molecular markers in the detection of colorectal adenoma, we added 50 colorectal adenoma samples to further screen the 10 CTCF binding sites screened in Example 1 Adding the mass spectrometry data of 40 samples in Example 1, this step of screening included a total of 90 samples, of which 20 were normal controls, 57 were adenomas, 6 were stage I tumors and 7 were stage II tumors. Table 6 shows the performance of these 10 CTCF binding sites in distinguishing tumor tissue from normal tissue. It can be seen from Table 6 that these 10 CTCF binding sites all have a strong ability to distinguish tumor tissue from normal tissue (AUC ≥ 0.85). Their sensitivities ranged from 44.64% to 88.89% at a specificity of 95%. Our goal is to establish an early diagnosis technology for colorectal tumors that includes as few molecular markers as ...

Embodiment 3

[0061] Example 3: Verification of the detection accuracy of the CTCF binding site CTCF_13 of the present application in a large sample

[0062] In order to verify the detection accuracy of the binding site CTCF_13 to be protected in this application, we selected 379 colorectal samples (Table 1), including 84 normal samples and 295 tumor samples (108 adenomas, 39 stage I tumors) , 101 cases of stage II tumors and 47 cases of stage III tumors), continue to use DNA mass spectrometry to detect the DNA methylation level of the binding site in 379 cases of colorectal samples, the detection method is the same as in Example 1 "DNA mass spectrometry DNA methylation analysis" has the same content, using the primer pair shown in SEQ ID NO:2 and SEQ ID NO:3.

[0063] According to the results of DNA mass spectrometry, we statistically analyzed the sensitivity of CTCF_13 in the detection of adenoma, stage I tumor, stage II tumor, stage III tumor and all tumors when the AUC value and specifi...

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Abstract

The invention discloses a novel use of a DNA (deoxyribonucleic acid) binding site CTCF_13 of a multifunctional transcription regulation factor CTCF, i.e. application thereof to preparation of a kit for early diagnosis of colorectal neoplasms. The nucleotide sequence of the binding site is shown as SEQ ID NO:1. The accuracy of the binding site CTCF, provided by the invention, in detection of either colorectal adenomas or the colorectal neoplasms is remarkably higher than that of a reported DNA methylated molecular marker; moreover, the DNA binding site of the CTCF is also likely to become a biomarker for tumor treatment effect monitoring, prognosis effect evaluation and tumor molecular typing (such as CIMP typing). In brief, a novel concept is provided for seeking for a more effective molecular marker for early diagnosis of tumors, treatment effect monitoring, prognosis effect evaluation and molecular typing, and is significant for tumor prevention and treatment.

Description

technical field [0001] The invention discloses the application of a DNA binding site CTCF_13 of a multifunctional transcription regulation factor CTCF. Background technique [0002] Colorectal cancer, one of the most morbid and fatal cancers in the world, results from the accumulation of genetic and epigenetic alterations in the colonic epithelium; approximately 1,235,108 people are diagnosed with colorectal cancer each year The number of deaths per year is about 609,051; the World Health Organization estimates that by 2030, the number of confirmed cases of colorectal cancer will increase by 77%, and the number of deaths will increase by 80%. The morbidity and death toll in China are increasing year by year, and the prevention and control situation of colorectal cancer is very severe. [0003] At present, most colorectal cancers develop from adenomas, and the time from adenomas to malignant tumors is about 10-15 years, which means that we have 10-15 years before turning int...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6886C12Q2600/112C12Q2600/118C12Q2600/154
Inventor 刘江唐莉鲁有望矣金华
Owner FIRST AFFILIATED HOSPITAL OF KUNMING MEDICAL UNIV
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