Primers, kits, detection methods and applications for detecting mir-874-3p expression

A kit and drug technology, applied in biochemical equipment and methods, drug combinations, pharmaceutical formulations, etc., can solve the problems of chemotherapy failure, tumor cell drug resistance, etc., and achieve the goal of improving chemotherapy effect, improving curative effect and reducing side effects Effect

Active Publication Date: 2020-11-20
优联瑞康(上海)基因科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Vincristine (vincristine, VCR) is currently a widely used anti-tumor chemotherapy drug for the clinical treatment of colon cancer, but long-term treatment will cause tumor cells to develop drug resistance, resulting in the failure of chemotherapy

Method used

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  • Primers, kits, detection methods and applications for detecting mir-874-3p expression
  • Primers, kits, detection methods and applications for detecting mir-874-3p expression
  • Primers, kits, detection methods and applications for detecting mir-874-3p expression

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] The primers used to detect the expression of miR-874-3p were designed and synthesized according to the sequence of miR-874-3p (as shown in SEQ ID NO: 1), including reverse transcription primers and fluorescent quantitative PCR primers, and the reverse transcription primers were as shown in SEQ ID NO: As shown in 2, 3 or 4, the forward primer in the fluorescent quantitative PCR primer is shown in SEQ ID NO: 5 or 6, and the reverse primer is shown in SEQ ID NO: 7.

Embodiment 2

[0040] The kit for detecting the expression of miR-874-3p, including the primers in Example 1 and the reverse transcription primer of the internal reference U6 snRNA (as shown in SEQ ID NO: 10) and fluorescent quantitative PCR primers (forward and reverse primers are as shown in SEQ ID NO: 10). ID NO: 8, 9), also includes general DNA digestion reagents (DNase I (RNase-free), EDPC, etc.), miRNA reverse transcription reagents (reverse transcriptase, 5×RT buffer, dNTP, etc.) and fluorescence quantification Polymerase chain reaction reagents (dNTP, 2×SYBR Green PCRMaster Mix, etc.).

Embodiment 3

[0042] A method for detecting the expression of miR-874-3p, comprising the following steps:

[0043] 1) Primer design

[0044] Primers were designed according to the mature sequence of hsa-miR-874-3p (No: MIMAT0004911) in the miRBase database (http: / / www.mirbase.org);

[0045]hsa-miR-874-3p: 5′-CUGCCCUGGCCCGAGGGACCGA-3′;

[0046] Reverse transcription primers:

[0047] Reverse transcription primer 1: 5'-CTCAACTGGTGTCGTGGAGTCGGCAATTCTCGGTCCC-3';

[0048] Reverse transcription primer 2: 5'-CTCAACTGGTGTCGTGGAGTCGGCAATTCTCGGTCC-3';

[0049] Reverse transcription primer 3: 5'-CTCAACTGGTGTCGTGGAGTCGGCAATTCTCGGTC-3';

[0050] Fluorescence quantitative PCR primers:

[0051] Forward primer 1: 5'-ACACTCCAGCTGGGCTGCCCTGGCCCGAGGG-3';

[0052] Forward primer 2: 5'-ACACTCCAGCTGGGCTGCCCTGGCCCGAGG-3';

[0053] Reverse primer: 5'-CTCAACTGGTGTCGTGGAGT-3';

[0054] The following primers were designed according to the U6 snRNA housekeeping gene (Genbank: NR_004394.1) in the NCBI database ...

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Abstract

The invention discloses a primer, a kit, a detection method and an application for detecting the expression of miR-874-3p, and belongs to the technical field of tumor biotherapy. The present invention proves that miR-874-3p is related to vincristine drug resistance through experiments, and is highly expressed in vincristine drug-resistant cells and lowly expressed in vincristine drug-resistant cells, so it can be used as a marker of tumor cell vincristine drug resistance. The primers, kits and detection methods designed based on the marker can be used to clinically judge the drug resistance of tumor cells to vincristine, further guide clinical medication and achieve precise treatment.

Description

technical field [0001] The invention relates to a primer for detecting the expression of miR-874-3p, a kit and a detection method comprising the primer, and the application of miR-874-3p in the preparation of antagonizing tumor vincristine-resistant drugs, It belongs to the field of tumor biotherapy technology. Background technique [0002] Colorectal cancer is one of the most common malignant tumors of the digestive tract, which occurs in the colon, preferably at the junction of the rectum and the sigmoid colon. Colorectal cancer has the highest incidence in the age group of 40 to 50 years old, with a male to female ratio of 2 to 3:1. In western developed countries, the incidence of colorectal cancer ranks third. In my country, the incidence of colorectal cancer ranks fourth, and the incidence and mortality are increasing year by year. Clinical data show that the high metastasis rate, low early diagnosis rate and easy recurrence after surgical resection of colorectal can...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886C12N15/11A61K45/06A61K31/475A61P35/00
CPCA61K31/475A61K45/06C12Q1/6886C12Q2600/106C12Q2600/136C12Q2600/158C12Q2600/178
Inventor 王天云董卫华高建辉林艳郭潇王燕芳
Owner 优联瑞康(上海)基因科技有限公司
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