MicroRNA marker group and its application in the preparation and evaluation of breast cancer chemosensitivity kit

A kit and sensitive technology, applied in the field of genetic testing, can solve the problems of difficulty in screening the main evaluation markers and difficult evaluation of the effect, and achieve the effect of large development value, improved accuracy, and important clinical significance.

Active Publication Date: 2019-10-22
SHANDONG JUNTENG MEDICAL TECH CO LTD
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  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The main reason is that the principle of tumor drug resistance has not been fully studied, so it is difficult to screen the main evaluation markers, and it is not easy to observe and evaluate the sensitivity of breast cancer cells to chemotherapy due to the tumor samples before and after breast cancer chemotherapy , making it difficult to evaluate the effects of markers after screening

Method used

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  • MicroRNA marker group and its application in the preparation and evaluation of breast cancer chemosensitivity kit
  • MicroRNA marker group and its application in the preparation and evaluation of breast cancer chemosensitivity kit
  • MicroRNA marker group and its application in the preparation and evaluation of breast cancer chemosensitivity kit

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Experimental program
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Effect test

Embodiment 1

[0031] 1. From 2009 to 2013, a total of 190 breast cancer tissue samples (breast cancer puncture specimens before chemotherapy and breast cancer surgical resection specimens after chemotherapy, in a paired relationship), of which 119 were from Qilu Hospital of Shandong University (randomly divided into two groups: primary screening and verification Sample), in order to exclude regional differences, another 71 cases of breast cancer patients from Liaocheng People's Hospital. According to the effect of neoadjuvant chemotherapy, 190 patients were graded according to the MP (Miller Payne) treatment response evaluation. Among them, the effect of chemotherapy in grade 1 and grade 2 was poor (that is, there was no response to chemotherapy or the reduction of tumor cells after chemotherapy was less than 30%), which was called chemotherapy. Drug-resistant group; grades 3, 4, and 5 have better chemotherapy effects (tumor cells are reduced by 30-90% after chemotherapy, and more than 90% a...

Embodiment 2

[0043] 1. Extraction of miRNA in breast cancer specimen tissue: Bioteke company's miRNA extraction kit in paraffin-embedded tissue.

[0044] (1) Tissue section: Cut the wax block of the breast cancer specimen into 10 slices of 4 μm;

[0045] (2) Dewaxing and dehydration of slices: put the tissue slices into a 65°C incubator for 1 hour to melt the wax, and then soak in 75%, 85%, 95% and 100% ethanol in xylene;

[0046] (3) Section nuclear staining: place the section in the newly prepared hematoxylin solution for 40-60s, gently rinse and dry;

[0047] (4) Tissue picking: Operate a 1ml syringe under a microscope to gently scrape off the cancer tissue in the section with its needle tip, and place it in a mixture of 240 μl PTL solution and 10 μl proteinase K;

[0048] (5) miRNA extraction:

[0049] 1) Water bath at 55°C and 80°C for 10 minutes each.

[0050] 2) Add 750 μl of Lysis Buffer MRL (purchased from Bioteke Company) to mix well, and let stand at room temperature for 3 mi...

Embodiment 3

[0073] Using GraphPad Prism 5 software, the receiver operating characteristic curve (receiver operating characteristic curve, ROC curve) analysis was performed on the specificity and sensitivity of the six miRNAs in distinguishing chemotherapy effects. In 60 primary screening cases, the results showed that miR-23a-3p, miR-200c-3p, miR-214-3p miR-638 and miR-451a (area under the curve AUC = 0.656, 0.727, 0.660, 0.658, 0.729, P All image 3 A; while the analysis results of miR-23b-3p showed that its AUC=0.638 (P>0.05), the difference was not statistically significant, such as Figure 4 shown.

[0074] Therefore, after removing miR-23b-3p, the remaining 5 indicators were used to construct the risk formula of chemotherapy resistance using the Logistic regression model:

[0075] Risk score=(0.782×expression of miR-23a-3p)-(3.968×expression of miR-638)+(0.361×expression of miR-200c-3p)+(5.848×expression of miR-214-3p)-(2.598 ×expression of miR-451a)+0.562.

[0076] According to thi...

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Abstract

The invention relates to a microRNA (micro Ribonucleic Acid) marker group and application thereof to preparation of a kit for evaluating the chemotherapy sensitivity of breast cancer. The microRNA marker group is composed of miR-638, miR-451a, miR-23a-3p, miR-214-3p and miR-200c-3p. A neoadjuvant chemotherapy clinical sample is used for carrying out microarray analysis for the first time and an inventor obtains the microRNA marker group with 5 miRNA markers through screening; and detection results of the 5 miRNA markers are compared with an established risk formula and whether the chemotherapy effect is good or not can be distinguished very well, so that the accuracy of predication is extremely improved.

Description

technical field [0001] The invention relates to a microRNA marker group and its application in the preparation of a breast cancer chemosensitivity kit, belonging to the technical field of gene detection. Background technique [0002] Breast cancer is one of the common malignant tumors in women, and its incidence rate accounts for 7-10% of all kinds of malignant tumors in the whole body. It is second only to uterine cancer in the incidence of malignant tumors in women. Gradually rising and rejuvenating trend, it has become one of the important killers that seriously threaten women's health. In recent years, neoadjuvant chemotherapy for breast cancer has been more and more widely used clinically. However, due to the high heterogeneity of tumors, tumors with the same histological type and TNM stage have different curative effects on the same treatment regimen. Effective chemotherapy can reduce tumor staging, increase surgical resection rate, eliminate micrometastases and reduc...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886C12N15/11
Inventor 高鹏邢爱艳王亚文李玉红陈旭
Owner SHANDONG JUNTENG MEDICAL TECH CO LTD
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