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A protein index for predicting the sensitivity of patients with esophageal squamous cell carcinoma to chemotherapy drugs and its application

A technology for esophageal squamous cell carcinoma and chemotherapeutic drugs, applied in the field of biomedicine, can solve problems such as insignificant correlation, and achieve the effect of shortening the treatment time

Active Publication Date: 2022-05-31
THE SECOND XIANGYA HOSPITAL OF CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, in vitro drug susceptibility tests have been conducted on patients for its drug resistance, but the correlation is not significant

Method used

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  • A protein index for predicting the sensitivity of patients with esophageal squamous cell carcinoma to chemotherapy drugs and its application
  • A protein index for predicting the sensitivity of patients with esophageal squamous cell carcinoma to chemotherapy drugs and its application
  • A protein index for predicting the sensitivity of patients with esophageal squamous cell carcinoma to chemotherapy drugs and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] Example 1 In Vitro Cell Experiment Study Regulating FBXO31 Affects Sensitivity of Esophageal Squamous Cell Carcinoma Cells to Paclitaxel

[0069] Paclitaxel, which is commonly used in clinic, is used as a drug, and the established stable FBXO31 silencing and control cell lines (refer to Liu, J., L. Han, B. Li, J. Yang, M.S. Huen, X. Pan, S. W. Tsao and A.L.Cheung(2014). "F-box only protein 31(FBXO31) negatively regulates p38 mitogen-activated protein kinase(MAPK) signaling by mediating lysine 48-linkedubiquitination and degradation of mitogen-activated protein kinase 6(MKK6)." J Biol Chems 289(31):21508-21518.) to detect the effect of FBXO31 gene on apoptosis.

[0070] Two kinds of cells (FBXO31 silenced cell line and control cell line) in the logarithmic growth phase were seeded in 6-well plates at a density of 0.5–1×10 4 Cells / well were treated with DMSO and paclitaxel (4 mg / ml) for 48 hours, and the apoptosis was detected.

[0071] Apoptosis detection methods i...

Embodiment 2

[0092] Example 2 Regulates the molecular biological mechanism of FBXO31

[0093] 1. Sample Preparation

[0094] (1) Materials and reagents

[0095]

[0096] (2) In-gel enzymatic hydrolysis

[0097] Gel strips were minced and destained using 50% acetonitrile containing 50 mM ammonium bicarbonate (NH4HCO3). The gel pieces were dehydrated by incubating with 100% acetonitrile for 5 minutes, and then the liquid phase in the system was removed, a dithiothreitol solution with a final concentration of 10 mM was added, and incubated at 37° C. for 60 minutes. Incubate with 100% acetonitrile again for dehydration, remove the liquid phase, add iodoacetamide (iodoacetamide) at a final concentration of 55 mM, and incubate at room temperature for 45 minutes in the dark. Afterwards, wash with ammonium bicarbonate at a final concentration of 50 mM, and incubate with 100% acetonitrile again for dehydration. Finally, the gel was resuspended in 50 mM ammonium bicarbonate containing 10 ng / μ...

Embodiment 3

[0108] Example 3 The activity of cofilin-1 can be regulated by regulating FBXO31 protein

[0109] On the first day, stable FBXO31 silenced cells and control cell lines were inoculated in six-well plates. On the second day, proteins were extracted after treatment with paclitaxel for 48 hours, and the expression levels of p-cofilin, cofilin and actin were detected by WB.

[0110] The results show Figure 5 : Knocking down FBXO31 protein can increase the phosphorylation level of cofilin-1, which means that the activity of cofilin-1 is reduced.

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Abstract

The invention belongs to the technical field of biomedicine, and in particular relates to a protein index for predicting the sensitivity of patients with esophageal squamous cell carcinoma to chemotherapeutic drugs and its application. The present invention finds that the up-regulation of FBXO31 protein expression in patients with esophageal squamous cell carcinoma can lead to a decrease in the phosphorylation level of cofilin-1 and an increase in the activity of cofilin-1, thereby causing drug resistance to paclitaxel in patients. Therefore, the up-regulation of FBXO31 protein expression may indicate that patients are resistant to the chemotherapy drug paclitaxel. Before treating patients with esophageal squamous cell carcinoma, detect the expression of FBXO31 protein in the patient's body by RT-PCR or immunohistochemistry to judge the patient's sensitivity to chemotherapy drugs, and provide a judgment index for doctors during the treatment process. The choice of therapeutic drugs has guiding significance, and the drug regimen can be adjusted in time, which shortens the treatment time for patients with esophageal squamous cell carcinoma to a certain extent.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a protein index for predicting the sensitivity of patients with esophageal squamous cell carcinoma to chemotherapeutic drugs and its application. Background technique [0002] Esophageal cancer is one of the digestive tract malignant tumors that seriously threaten human life and health. In my country, about 90% of esophageal cancers are squamous cell carcinomas of the esophagus. Chemotherapy is one of the important treatment methods for esophageal cancer. The use of chemotherapy drugs plays an important role in inhibiting tumor growth, recurrence and metastasis. However, primary multidrug resistance and secondary drug resistance are one of the important reasons leading to treatment failure and low survival rate of esophageal cancer. Commonly used anticancer drugs such as alkylating agents, platinum complexes, and topoisomerase inhibitors (using DNA as the target,...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68A61K31/337A61P35/00
CPCG01N33/6893A61K31/337A61P35/00G01N2800/52
Inventor 刘佳吕梁杨慧王树超李毅夫邓军卫
Owner THE SECOND XIANGYA HOSPITAL OF CENT SOUTH UNIV
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