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A biomarker for detecting the prognosis of nasopharyngeal carcinoma and its application

A biomarker and nasopharyngeal carcinoma technology, applied in the field of biomedicine, can solve the problems of large differences in patient prognosis, avoid over-treatment, relieve economic pressure and medical resource pressure, and improve prediction accuracy

Active Publication Date: 2022-05-17
AFFILIATED HOSPITAL OF NANTONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the clinicopathological stage of the patient is mainly used to judge the prognosis and formulate the treatment strategy. However, the prognosis of patients with the same clinicopathological stage is often quite different. Therefore, there is an urgent need for a more accurate prognosis of nasopharyngeal carcinoma. Methods

Method used

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  • A biomarker for detecting the prognosis of nasopharyngeal carcinoma and its application
  • A biomarker for detecting the prognosis of nasopharyngeal carcinoma and its application
  • A biomarker for detecting the prognosis of nasopharyngeal carcinoma and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1 (exploring the relationship between the biomarker miR-106a-5p and the prognosis of patients with nasopharyngeal carcinoma)

[0037] In order to explore the role of the biomarker miR-106a-5p in the progression of nasopharyngeal carcinoma, we performed an in situ hybridization (ISH) experiment on the above-mentioned 123 tissue samples, including the following steps:

[0038] Step 1: Dewax and hydrate the tissue sample chip (this step needs to be performed in a fume hood)

[0039] Soak the tissue sample chip in xylene for 30 minutes, and replace the xylene every 10 minutes; then fully soak the tissue sample chip soaked in xylene in 100% ethanol, soak in 100% ethanol for 5 minutes, and fully soak in 96% ethanol , soak in 96% ethanol for 5 minutes, fully soak in 70% ethanol, soak in 70% ethanol for 5 minutes, and soak in PBS for 5 minutes.

[0040] Step 2: Digest the tissue sample chip with protease

[0041] Submerge the dewaxed tissue sample chip in proteinase K...

Embodiment 2

[0057] Example 2 (demonstration that the biomarker BTG3 is a direct target of miR-106a-5p)

[0058] The biologically effective interaction between miR-106a-5p and BTG3 was confirmed by the luciferase reporter gene experiment. The specific operation steps of the luciferase reporter gene experiment are as follows:

[0059] Step 1: Transfection of plasmids into nasopharyngeal carcinoma CNE2 cells

[0060] 1. Experimental group:

[0061] Group A: Plasmids are 106NC and PGL3-Mut

[0062] Group B: Plasmids are 106NC and PGL3-WT

[0063] Group C: Plasmids are 106Mimics and PGL3-Mut

[0064] Group D: Plasmids 106Mimics and PGL3-WT

[0065] 2. Inoculate CNE2 cells into a six-well plate, set 3 replicates in each group, and then inject medium into the six-well plate for cell culture. When the cell density reaches 40%-50%, add transfection reagents and corresponding plasmids for co-transfection.

[0066] Co-transfection process: Add 5 μL of Lip3000 transfection reagent to 1 mL of ba...

Embodiment 3

[0072] Example 3 (the relationship between the biomarker BTG3 and the prognosis of patients with nasopharyngeal carcinoma)

[0073] experimental method:

[0074] In order to explore the role of the biomarker BTG3 (i.e. miR-106a-5p target gene) in the progression of nasopharyngeal carcinoma, immunohistochemical experiments were also performed on 123 tissue samples for BTG3, including the following steps:

[0075] Step 1: Dewax and hydrate the tissue sample chip (this step needs to be performed in a fume hood)

[0076] Soak the tissue sample chip in xylene for 30 minutes, and replace the xylene every 10 minutes; then fully soak the tissue sample chip soaked in xylene in 100% ethanol, soak in 100% ethanol for 5 minutes, and fully soak in 96% ethanol , soak in 96% ethanol for 5 minutes, fully soak in 70% ethanol, soak in 70% ethanol for 5 minutes, and soak in PBS for 5 minutes.

[0077] Step 2: Antigen retrieval, quenching, blocking

[0078] 1. Heat 0.01 sodium citrate buffer s...

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Abstract

The invention belongs to the field of biomedicine and discloses a biomarker for detecting the prognosis of nasopharyngeal carcinoma and its application. The biomarker proposed by the present invention consists of one or three of miR-106a-5p, BTG3 and SOX9. When the biomarkers consisted of miR‑106a‑5p, BTG3 and SOX9, the score based on the quantitative expression of the three biomarkers could rapidly and accurately predict the prognosis of nasopharyngeal carcinoma. If this combination of biomarkers is applied to the preparation of reagents or tools for detecting the prognosis of nasopharyngeal carcinoma, it will significantly improve the prediction accuracy of the prognosis of nasopharyngeal carcinoma patients, and then screen a large number of nasopharyngeal carcinoma patients with poor prognosis. Group, while ensuring that these patients receive reasonable treatment, it also avoids overtreatment of patients with better prognosis, relieves the economic pressure of patients and society and pressure on medical resources, and is conducive to reducing treatment costs and improving the quality of life of patients with nasopharyngeal carcinoma. prognosis.

Description

technical field [0001] The invention belongs to the field of biomedicine, in particular to a biomarker for detecting the prognosis of nasopharyngeal carcinoma and its application. Background technique [0002] Nasopharyngeal carcinoma (NPC) is the most common malignant tumor of head and neck cancer. The incidence rate in China is the highest in the world, accounting for about 80% of the world, and the incidence rate is 15-30 per 100,000 people. At present, there are many methods for the treatment of nasopharyngeal carcinoma, such as radiotherapy, chemotherapy combined with radiotherapy and chemotherapy, targeted drugs, etc., which need to be selected reasonably according to the actual situation of the patient. What is the prognosis of each nasopharyngeal carcinoma patient, and how to make different diagnosis and treatment plans according to their possible different prognosis so as to get better treatment effect is one of the most concerned issues in clinical practice. At pr...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886C12N15/113
CPCC12Q1/6886C12Q2600/118C12Q2600/158C12Q2600/178
Inventor 尤易文游波张启成顾苗夏天陈静张薇张洁吴笛施思单颖包丽丽
Owner AFFILIATED HOSPITAL OF NANTONG UNIV
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