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Application of substance for inhibiting expression of TGF-betaR1 in treatment and/or prevention of esophageal cancer

A TGF-, esophageal cancer technology, applied in the field of biomedicine, can solve the problems of lost optimal timing, complex pathological changes of esophageal cancer, and failure of chemotherapy

Pending Publication Date: 2021-05-28
SHIHEZI UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the lack of specific and sensitive early diagnostic indicators and methods, nearly half of the patients with esophageal cancer have metastasized at the time of diagnosis, thus losing the best opportunity for surgical resection of all lesions. Drug resistance is the main cause of chemotherapy failure
Although scientists have conducted in-depth research on genetic and environmental factors in recent decades, the pathogenesis of esophageal cancer has not been fully clarified due to the complex pathological changes of esophageal cancer. important for treatment and prognosis

Method used

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  • Application of substance for inhibiting expression of TGF-betaR1 in treatment and/or prevention of esophageal cancer
  • Application of substance for inhibiting expression of TGF-betaR1 in treatment and/or prevention of esophageal cancer
  • Application of substance for inhibiting expression of TGF-betaR1 in treatment and/or prevention of esophageal cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0070] Example 1. High expression of TGF-βR1 in esophageal squamous cell carcinoma cells and esophageal cancer stem cell spheres after co-culture

[0071] Enrichment of esophageal cancer stem cells by natural screening method in serum-free culture and detection of OCT-4, CD44 and P75 by qRT-PCR NTR The expression of genes related to stem cell characteristics was different in esophageal cancer stem cell spheres (experimental group) and EC109 or EC9706 adherent cells (control group), and the expression of TGF-βR1 in stem cell spheres was detected by Western blot.

[0072] Esophageal cancer stem cell spheres: Inoculate 10,000 esophageal cancer cell lines EC109 or EC9706 in the logarithmic growth phase into an ultra-low adhesion six-well plate containing serum-free stem cell medium (F12 serum-free medium) , after the sphere cells are formed, the tumor stem cell spheres are collected.

[0073] Co-cultivation with M2 macrophages: The esophageal cancer cell line EC109 or EC9706 in t...

Embodiment 2

[0080] Example 2. Preparation of Esophageal Cancer Cells with Reduced Expression of TGF-βR1 and Its Application in Inhibiting Stem Cell Characteristics, Proliferation, Migration and Invasion of Esophageal Cancer Cells

[0081] 1. SB431542 can reduce the expression of TGF-βR1 in esophageal squamous cell carcinoma cells

[0082] Since the inhibitor usually has a certain degree of cytotoxicity, in order to reduce its cytotoxic effect as much as possible, the inventors used multiple time points such as 0h, 12h, 24h, 36h, 48h to detect the TGF-βR1 inhibitor by using the CCK8 method (ie SB431542) the change of cell proliferation ability after treatment, the cells to be tested are EC109 cells or EC9706 cells.

[0083] The specific steps for detecting cell proliferation ability by CCK8 method are as follows: culture the cells to be tested in the cell culture well plate, when the cell content is 10000, add TGF-βR1 inhibitor to the culture, the concentration of TGF-βR1 inhibitor is set ...

Embodiment 3

[0116] Example 3, the role of overexpression of TGF-β1 in promoting the stemness of esophageal cancer cells, and further promoting the proliferation, migration and invasion of stem cells.

[0117] 1. The application of substances overexpressing TGF-β1 in inhibiting the stemness of esophageal cancer cells

[0118] In order to detect the effect of adding the TGF-β1 recombinant agent rhTGF-β1 (product number 100-21, Peprotech (UK)) on the stemness of esophageal cancer, Western Blot, qRT-PCR and sphere culture were used to detect the effect of adding TGF-β1. Changes in the expression of stemness markers and sphere formation rate of EC109 and EC9706 cells after β1 recombinant agent, and the negative control cells (ie, EC109 or EC9706 cells) were used as the control group. Proceed as follows:

[0119]Esophageal cancer cell lines EC109 or EC9706 in the logarithmic growth phase were inoculated at 10,000 cells / well in an ultra-low adhesion six-well plate containing serum-free stem cel...

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Abstract

The invention discloses application of a substance for inhibiting TGF-betaR1 expression in treatment and / or prevention of esophageal cancer. It finds that TGF-betaR1 is highly expressed in esophageal cancer stem cell spheres and can promote proliferation and self-renewal of esophageal squamous carcinoma cells and formation, migration and invasion of tumor stem cell spheres, and substances for inhibiting expression of TGF-betaR1 can reverse the phenomenon. The invention further provides a theoretical basis and an experimental basis for defining the occurrence and development mechanism of esophageal cancer and targeted therapy.

Description

technical field [0001] The invention relates to the application of substances inhibiting the expression of TGF-βR1 in the treatment and / or prevention of esophageal cancer in the field of biomedicine. Background technique [0002] Esophageal cancer is the sixth leading cause of cancer-related deaths. Most of esophageal cancer occurs in developing countries, especially in China, where the incidence of esophageal cancer is about 70% of the total number of esophageal cancer in the world. The five-year survival rate for esophageal cancer is only about 15%, and most patients die within a year of diagnosis. Due to the lack of specific and sensitive early diagnostic indicators and methods, nearly half of the patients with esophageal cancer have metastasized at the time of diagnosis, thus losing the best opportunity for surgical resection of all lesions. Drug resistance is the main reason for the failure of chemotherapy. Although scientists have conducted in-depth research on gene...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K45/00A61K31/4439A61P1/00A61P35/00
CPCA61K45/00A61K31/4439A61P1/00A61P35/00
Inventor 胡建明李疆芬李玲李锋江辰昊
Owner SHIHEZI UNIVERSITY
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