Application of specific methylation sites as breast cancer molecular subtype diagnosing markers

A molecular typing and methylation technology, applied in the determination/examination of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., and can solve problems such as non-minimally invasive or non-invasive breast cancer

Inactive Publication Date: 2018-10-19
BEIJING INST OF GENOMICS CHINESE ACAD OF SCI CHINA NAT CENT FOR BIOINFORMATION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The present invention aims to provide the application of a specific methylation site as a diagnostic marker for molecular typing of breast cancer

Method used

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  • Application of specific methylation sites as breast cancer molecular subtype diagnosing markers
  • Application of specific methylation sites as breast cancer molecular subtype diagnosing markers
  • Application of specific methylation sites as breast cancer molecular subtype diagnosing markers

Examples

Experimental program
Comparison scheme
Effect test

Embodiment approach

[0078] According to a typical embodiment of the present invention, the device further includes an evaluation module, which is used to compare the detection value of the subject with the detection value of breast cancer patients with different molecular types, when selected from chr10: 39097581, chr10: 88386316, chr11: 105772483,chr11:113528661,chr1:163455492,chr1:164069493,chr1:173149222,chr1: 201310696,chr12:29221374,chr1:233270329,chr1:242355017,chr14:60666657,chr1: 59426963,chr16:46420219,chr18: 61618871,chr21:20247091,chr21:23757716,chr2: 36975610,chr2:87314115,chr2:89834041,chr2:89837315,chr2:89858946,chr3:115818550, chr3:144975142,chr4:164139191,chr4:189484234,chr4:3579745, chr4:47297495,chr4: 71802931,chr5:42079715,chr6:100477543,chr6:153594933,chr6:40877403,chr6:75723365, chr7:103135406,chr7:148667863,chr7:43817571,chr8:27578331,chr8:46896342和chr8: The methylation changes of one or more sites in 75989278 are in the same trend compared with the detected values ​​of pati...

Embodiment 1

[0098] Collection and identification of blood samples from breast cancer patients

[0099] Blood samples from patients with different molecular types were collected and collected through verification by experienced physicians (the samples were provided by Tianjin Cancer Hospital).

Embodiment 2

[0101] Extraction and library sequencing of cfDNA in breast cancer patients

[0102] Blood samples from patients with various breast cancer molecular types were collected, and cfDNA was extracted from plasma. Utilize the QIAamp Circulating Nucleic Acid Kit.

[0103] The specific operation steps are as follows: 1) Use the density gradient centrifugation method to centrifuge at a speed of 1900g to separate red blood cells, white blood cells and plasma. The clear top layer is plasma, take out the plasma, and then use a speed of 14000rpm to perform high-speed centrifugation again. The upper layer is plasma; 2) Add the extracted plasma from 1) to a 50ml centrifuge tube that has been added with proteinase K, and then add the lysate ACL (which already contains CarrierRNA) to obtain the first mixed solution; 3) Add the last obtained second mixture in 2) 1. Mix the mixed solution and put it in a water bath at 60°C for 30 minutes; 4) Take out the first mixed solution from the water bat...

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Abstract

The invention discloses application of specific methylation sites as breast cancer molecular subtype diagnosing markers. The invention establishes a method and a standard for screening disease-relatedchromosomal methylation sites, and screens 40 methylation sites related to breast cancer diagnosis by using breast cancer patients as cases. The screened specific methylation sites as the breast cancer molecular subtype diagnosing markers can be applied to non-invasive detection of molecular breast cancer subtypes and diagnosis of other diseases. A kit is used for one or more of the following applications: diagnosis of the breast cancer molecular subtypes, assessment of breast cancer risks, assessment of a breast cancer treating effect, screening of breast cancer treating medicines and the like.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the application of a specific methylation site as a diagnostic marker for molecular typing of breast cancer. Background technique [0002] Liquid biopsy technology was rated as one of the top ten scientific and technological advances in 2015 by MIT technology, providing a new idea for the study of tumor drug resistance biomarkers. Compared with tissue biopsy, liquid biopsy has many advantages, such as non-invasive and reproducible tumor samples, less side effects, simple operation, lower cost, and faster detection speed. [0003] Circulating tumor DNA (cfDNA) carries the mutation and methylation information of tumor DNA. Using cfDNA as a tumor marker has low false positive rate, high sensitivity and high accuracy. Most protein tumor markers can exist in the blood for several weeks, and the half-life of cfDNA is less than two hours, so cfDNA can be used to monitor the development of ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886C12N15/11
CPCC12Q1/6886C12Q2600/154
Inventor 孙英丽安珂李国超
Owner BEIJING INST OF GENOMICS CHINESE ACAD OF SCI CHINA NAT CENT FOR BIOINFORMATION
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