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Application of trim50 in the diagnosis and treatment of esophageal squamous cell carcinoma

A technology of esophageal squamous cell carcinoma and reagents, applied in the field of biomedicine, can solve the problems of inability to block signal transduction and unsatisfactory treatment effect

Active Publication Date: 2019-06-11
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, most molecularly targeted drugs can only block one receptor, but not all signal transduction, and the therapeutic effect is often unsatisfactory

Method used

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  • Application of trim50 in the diagnosis and treatment of esophageal squamous cell carcinoma
  • Application of trim50 in the diagnosis and treatment of esophageal squamous cell carcinoma
  • Application of trim50 in the diagnosis and treatment of esophageal squamous cell carcinoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] Example 1 Screening for gene markers associated with esophageal squamous cell carcinoma

[0060] 1. Sample collection

[0061] Six cases of surrounding esophageal squamous cell carcinoma tissues and surrounding normal esophageal mucosal tissues were collected, and the patients gave informed consent. All the above specimens were obtained with the consent of the organizational ethics committee.

[0062] 2. Preparation of RNA samples (operated using QIAGEN tissue RNA extraction kit)

[0063] Take out the tissue samples frozen in liquid nitrogen, put the tissue samples into a pre-cooled mortar for grinding, and extract and isolate RNA according to the instructions in the kit. details as follows:

[0064] 1) Add Trizol and place at room temperature for 5 minutes;

[0065] 2) Add 0.2ml of chloroform, vibrate the centrifuge tube vigorously, mix well, and place it at room temperature for 5-10min;

[0066] 3) Centrifuge at 12000rpm for 15min, transfer the upper aqueous phase...

Embodiment 2

[0078] Example 2 QPCR sequencing to verify differential expression of TRIM50 gene

[0079] 1. Large-sample QPCR verification of differential expression of TRIM50 gene. According to the sample collection method in Example 1, 50 cases of surrounding normal esophageal mucosa tissues and 50 cases of esophageal squamous cell carcinoma tissues were selected.

[0080] 2. RNA extraction The steps are the same as in Example 1.

[0081] 3. Reverse transcription:

[0082] Use 25μl reaction system, take 1μg total RNA for each sample as template RNA, and add the following components to PCR tubes: DEPC water, 5× reverse transcription buffer, 10mM dNTP, 0.1mM DTT, 30μM Oligo dT, 200U / μl M-MLV, template RNA. Incubate at 42°C for 1h, then centrifuge briefly at 72°C for 10min.

[0083] (3) QPCR amplification test

[0084] Primer design

[0085] The primer sequence of TRIM50 gene is:

[0086] Forward primer: 5'-TGAAGCAGGAGCAGAAAA-3' (SEQ ID NO.3)

[0087] Reverse primer: 5'-GAAGACATCCGAC...

Embodiment 3

[0099] Example 3 Differential expression of TRIM50 gene in esophageal cancer cell lines

[0100] 1. Cell culture

[0101] Human esophageal squamous cell lines KYSE 150 and KYSE450 were purchased from Cancer Institute, Chinese Academy of Sciences, and normal esophageal epithelial cell line Het-1a was purchased from Guangzhou Geneo Company. In DMEM containing 10% fetal bovine serum and 1% P / S at 37°C, 5% CO 2 , Cultivated in an incubator with a relative humidity of 90%. Change the medium once every 2-3 days, and use 0.25% EDTA-containing trypsin for routine digestion and passage.

[0102] 2. Extraction of RNA

[0103] 1) Digest adherent cells with trypsin, and the cells obtained by pipetting are centrifuged, resuspended, washed, and then resuspended in 1640 medium (10% calf serum).

[0104] 2) Transfer the resuspended cells to a 6-well plate ( / well), add medium to 2ml / well, shake the 6-well plate gently to resuspend the cells evenly.

[0105] 3) The cells were grown on the ...

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Abstract

The invention discloses an application of TRIM50 in the diagnosis and treatment of esophageal squamous cell carcinoma. In the invention, the chip hybridization result is analyzed by a bioinformatics method; the expression of differential expression gene TRIM50 of esophageal squamous cell carcinoma is down-regulated in a patient; and by up-regulating the expression level of TRIM50, the proliferation of esophageal squamous cell carcinoma cells can be inhibited, and the apoptosis rate of esophageal squamous cell carcinoma cells is increased. The invention provides a theoretical basis for precision medicine as well as a new target for the research and development of drugs.

Description

technical field [0001] The invention belongs to the field of biomedicine, and relates to the application of TRIM50 in the diagnosis and treatment of esophageal squamous cell carcinoma. Background technique [0002] Esophageal cancer, also known as esophageal cancer, is one of the most common malignant tumors of the digestive tract in clinical practice. According to a report from the International Agency for Research on Cancer (IARC) of the World Health Organization, the incidence of esophageal cancer ranks eighth in the list of malignant tumors in the world, and its death rate ranks sixth. According to statistics, in 2008, the annual number of new cases of esophageal cancer was about 482,300, and the number of deaths was 406,800. The new cases and mortality rate of esophageal cancer in China rank first in the world. Pathological studies have proved that squamous cell carcinoma derived from squamous epithelium is the most common pathological type of esophageal cancer, while...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886G01N33/574A61K45/00A61P35/00
CPCA61K45/00C12Q1/6886C12Q2600/158G01N33/57407G01N33/57484
Inventor 杨承刚任静
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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