Biomarker for diagnosing and predicting breast cancer and detection kit

A breast cancer and reagent kit technology, applied in the field of tumor diagnosis, can solve problems that have not been reported yet

Inactive Publication Date: 2016-02-24
杨廷稳
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] So far, there have been no reports on the relationship between CystatinSN and CystatinS in the cysteine ​​protease inhibitor family and breast cancer

Method used

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  • Biomarker for diagnosing and predicting breast cancer and detection kit
  • Biomarker for diagnosing and predicting breast cancer and detection kit
  • Biomarker for diagnosing and predicting breast cancer and detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1: Magnetic bead method specifically enriches CystatinSNmRNA, CystatinSmRNA and RPN1mRNA in urine

[0024] According to the mRNA sequence design of CystatinSN, CystatinS and RPN1 (ribosome binding protein 1), the specific probe (the nucleotide sequence of CystatinSNmRNA such as SEQIDNO.1, the nucleotide sequence of CystatinSmRNA such as SEQIDNO.2, The nucleotide sequence of RPN1mRNA is as SEQIDNO.3): the probe for capturing CystatinSNmRNA is 5'-aaagagcacaactgtttcttctgca(dA)30-3'(SEQIDNO.4), the probe for capturing CystatinSmRNA is 5'-taccaggtctattagaagca(dA)30- 3' (SEQ ID NO.5), the probe for capturing the internal reference gene RPN1 mRNA is 5'-gatgagcttctcattctcaatgtacg(dA)30-3' (SEQ ID NO.6). The above-mentioned specific probes can complementarily bind to olig(dT) of magnetic beads (GE, catalog number: 3815-2103-010150) to obtain magnetic beads bound to specific probes.

[0025] The enrichment steps are as follows: fresh urine and sample transport preservati...

Embodiment 2

[0037] Example 2: Detection of CystatinSNmRNA and CystatinSmRNA expression in breast cancer tissue

[0038] 30 cases of breast cancer and adjacent paired tissue samples were collected from the Department of Medical Oncology, Jiangsu Cancer Hospital, cut to the size of rice grains, stored in RNAlater preservation solution at -80°C, and equilibrated to room temperature before use. Then according to the method of Example 1, the CystatinSNmRNA, CystatinSmRNA and RPN1mRNA of breast cancer and adjacent paired tissues were enriched respectively, and then respectively with CystatinSNmRNA detection primers, CystatinSmRNA detection primers and internal reference gene RPN1mRNA detection primers to detect 30 cases of sample breast cancer and adjacent cancerous pairings The relative expression levels of CystatinSNmRNA and CystatinSmRNA in tissues, the detection system and detection conditions are the same as those in Example 1. The test results are 2 -△△CP Calculate the relative expressio...

Embodiment 3

[0039] Example 3: Construction of breast cancer detection kit

[0040] 1. Construction of CystatinSN recombinant plasmid

[0041] The breast cancer cell line MCF-7 was used as the material to extract the total mRNA of the breast cancer cell line, and then the extracted mRNA was used as a template to synthesize cDNA, and the primers for constructing the CystatinSN recombinant plasmid were designed according to the sequence of the CystatinSN gene (or CST1 gene), and the CystatinSN recombinant plasmid The upstream primer is 5'-ctggagccccaaggagga-3' (SEQ ID NO.13), the downstream primer is 5'-accagtccagggggtggga-3' (SEQ ID NO.14), and the nucleotides shown in SEQ ID NO.13 and SEQ ID NO.14 are used as primers to synthesize The cDNA was used as a template for PCR amplification. The amplification conditions were: pre-denaturation at 94°C for 5 minutes; 45 cycles of denaturation at 94°C for 30 seconds, annealing at 60°C for 30 seconds, and extension at 72°C for 1 minute; extension at ...

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Abstract

The invention discloses a biomarker for diagnosing and predicting breast cancer and a detection kit. Multiple tumor markers are applied in a combined manner to diagnose and predict the breast cancer, and both specificity and sensitivity are obviously higher than those when a single tumor marker is used for diagnosing and predicting the breast cancer. The invention further provides a biological detection kit based on multiple markers. The kit is convenient to use, high in specificity and sensitivity, suitable for being used for diagnosis of the breast cancer, capable of realizing curative effect evaluation and monitoring of post-treatment metastasis and recurrence in the process of treatment and quite wide in clinical application prospect.

Description

technical field [0001] The invention belongs to the field of tumor diagnosis, and in particular relates to a breast cancer detection and diagnosis marker and a biological detection kit, which are used for auxiliary diagnosis and prediction of breast cancer. Background technique [0002] In 2012, CellResearch magazine published online the latest research results of Differential secretomeanalysisrevealsCST6asasuppressorofbreastcancerbonemetastasisby the research group of Hu Guohong, Institute of Health Sciences, Chinese Academy of Sciences, and the research group of Xu Xuemin and Hu Xiaofang, Shanghai Jiaotong University Med-X Center. The findings reveal that CST6 plays an important role in bone metastasis of breast cancer. [0003] Breast cancer is a malignant disease that poses the greatest threat to women's health. It involves a variety of genetic abnormalities, including the activation of oncogenes and the inactivation of tumor suppressor genes. Its development is a multi-...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6886C12Q2600/178
Inventor 杨廷稳
Owner 杨廷稳
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