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Bladder cancer marker lncRNA TERC based on urine exosomes and application thereof

An exosome and bladder cancer technology, applied in the field of bladder cancer marker lncRNATERC, can solve the problems of bladder cancer markers that have not been reported, and achieve excellent diagnostic performance, non-invasive and easy access, and high patient compliance

Pending Publication Date: 2021-11-12
SHANGHAI TONGJI HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are no reports about the urinary exosome-based bladder cancer marker lncRNA TERC and its application in the present invention.

Method used

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  • Bladder cancer marker lncRNA TERC based on urine exosomes and application thereof
  • Bladder cancer marker lncRNA TERC based on urine exosomes and application thereof
  • Bladder cancer marker lncRNA TERC based on urine exosomes and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1 Screening and extraction of exosome target molecule lncRNA—TERC

[0028] 1. In the present invention, exosomes are firstly extracted from the urine samples of 4 patients diagnosed with bladder cancer and 3 healthy individuals obtained from Tongji Hospital in Shanghai. The experimental flowchart of extracting urine exosomes by differential ultracentrifugation, see figure 1 .

[0029] The extracted exosomes are strictly identified by transmission electron microscopy (TEM), nanoparticle size tracking analysis (NTA) and western blot technology in accordance with ISEV2018 standards, and only qualified exosome samples can be used for downstream analysis.

[0030] 2. The specific steps for extracting exosomal RNA are as follows:

[0031] (1) Add 500 μl of anhydrous trizol to the exosome-PBS mixture, vortex and mix well, and then let stand at room temperature for 5 minutes to fully lyse;

[0032] (2) Add 100 μl chloroform, vortex and mix well, place at room tempera...

Embodiment 2

[0048] Example 2 Method for diagnosing bladder cancer based on urinary exosome lncRNA TERC

[0049] 1. Urine exosome extraction (PEG precipitation method)

[0050] (1) Use a 15ml sterile centrifuge tube to collect 8ml of the urine sample to be extracted, and store it at 4°C;

[0051] (2) Centrifuge the sample at 3000g for 15min at 4°C, discard the precipitate and collect the supernatant;

[0052] (3) Carefully transfer the supernatant into another clean 15ml centrifuge tube;

[0053] (4) Centrifuge the sample at 10,000 g for 20 min at 4°C, discard the precipitate and collect the supernatant;

[0054] (5) Carefully transfer the supernatant into another clean 15ml centrifuge tube;

[0055] (6) Add 2ml PEG6000 (ratio: 4:1) to 8ml of urine supernatant, close the cap of the centrifuge tube tightly, mix upside down for about 1min to make the liquid fully mixed, and evenly distribute the liquid into five 2mlEP tubes middle;

[0056] (7) Put it in a refrigerator at 4°C overnight ...

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Abstract

The invention relates to a bladder cancer marker lncRNA TERC based on urine exosomes and application thereof. The target molecule lncRNA TERC has high sensitivity and specificity in bladder cancer diagnosis and has excellent diagnosis performance, the blank of bladder cancer liquid biopsy diagnosis markers in clinic is filled, the sample type is urine, and the bladder cancer liquid biopsy diagnosis markers are noninvasive and easy to obtain, and the patient compliance is high. In general, the invention provides the bladder cancer liquid biopsy diagnosis marker with strong specificity and high sensitivity for clinic.

Description

technical field [0001] The present invention relates to the technical field of biomedicine, in particular to urinary exosome-based bladder cancer marker lncRNA TERC and its application. Background technique [0002] Bladder cancer (BLCA) is one of the common malignant tumors of the urinary system. With the aging of the population and the acceleration of industrialization, the incidence of bladder cancer is increasing year by year. According to statistics from the International Agency for Research on Cancer of the World Health Organization, the incidence of bladder cancer ranks tenth among all malignant tumors in 2020, and its incidence and mortality both rank second among malignant tumors of the male urinary system, second only to prostate cancer. Worldwide, the incidence of bladder cancer shows significant regional and gender differences. The global age-standardized incidence rates are 9.0 / 100,000 and 2.2 / 100,000 for men and women, respectively. Among EU countries, Belgium...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886C12N15/113
CPCC12Q1/6886C12Q2600/178C12Q2600/158
Inventor 李冬陈晨孙祖俊吴军录商安全
Owner SHANGHAI TONGJI HOSPITAL