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Application of CGA gene as target spot in preparation of drug for diagnosing and treating drug-resistant solid tumors

A technology based on solid tumors and genes, applied in drug combinations, antineoplastic drugs, biochemical equipment and methods, etc., can solve problems such as treatment failure

Pending Publication Date: 2022-02-15
FOURTH MILITARY MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Chemotherapy (chemotherapy) plays an important role in the treatment of solid tumors, however, the vast majority of patients undergo chemotherapy drug resistance leading to treatment failure

Method used

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  • Application of CGA gene as target spot in preparation of drug for diagnosing and treating drug-resistant solid tumors
  • Application of CGA gene as target spot in preparation of drug for diagnosing and treating drug-resistant solid tumors
  • Application of CGA gene as target spot in preparation of drug for diagnosing and treating drug-resistant solid tumors

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Example 1: Real-time quantitative PCR (Real-time qualitative PCR, RT-qPCR) detection of CGAmRNA in gastric cancer drug-resistant cell line SGC7901 ADR and SGC7901 VCR The expression difference between the cell line SGC7901 and the control group

[0054] 1. Research object

[0055] Gastric adenocarcinoma cell line SGC7901 is derived from the Academy of Military Medical Sciences and preserved by the State Key Laboratory of Tumor Biology, Air Force Military Medical University;

[0056] SGC7901 cells were cultured in DMEM medium containing 10% (v / v) fetal bovine serum;

[0057] Drug-resistant gastric cancer cell line SGC7901 ADR and SGC7901 VCR The cells were established by the intermittent induction method. The method was to expose SGC7901 parental cells to lethal doses of doxorubicin and vincristine for a short period of time and induce them successively. The alkali concentration is gradually increased from 1 μg / ml to 5 μg / ml; specifically, SGC7901 ADR For long-term c...

Embodiment 2

[0067] Example 2: WB detection of CGA protein in gastric cancer drug-resistant cell line SGC7901 ADR and SGC7901 VCR The expression difference between the cell line SGC7901 and the control group

[0068] 1. Research object

[0069] Drug-resistant gastric cancer cell line SGC7901 ADR and SGC7901 VCR The cultivation with the control group cell line SGC7901 was the same as in Example 1.

[0070] 2. Experimental method

[0071] Detection of CGA protein in SGC7901 by WB ADR and SGC7901 VCR and the expression level of SGC7901 cells, the specific steps are as follows:

[0072] After the normal culture of each cell for 24 hours, the DMEM medium without fetal bovine serum was replaced to continue the culture for 24 hours, and the culture supernatant and cells were collected respectively;

[0073] Use the Millipore ultrafiltration tube to centrifuge at 4°C and 4000×g for 1 h to collect the secreted protein in the culture supernatant, use the RIPA cell lysate to extract the total...

Embodiment 3

[0077] Example 3: Immunofluorescence (IF) detection of CGA protein in gastric cancer drug-resistant cell line SGC7901 ADR and SGC7901 VCR Differences in expression and subcellular localization between the cell line SGC7901 and the control group

[0078] 1. Research object

[0079] Drug-resistant gastric cancer cell line SGC7901 ADR and SGC7901 VCR The cultivation with the control group cell line SGC7901 was the same as in Example 1.

[0080] 2. Experimental method

[0081] Detection of CGA protein in SGC7901 by IF ADR , SGC7901 VCR and the expression and localization of SGC7901 cells, the specific steps are as follows:

[0082] After the cells were inoculated on chamber slides and cultured for 24 hours, 4% paraformaldehyde was added to fix the cells, 0.5% Triton was added, and the cells were permeabilized at room temperature for 15 minutes; goat serum blocking solution was added dropwise and left at room temperature for 30 minutes ; The primary antibody (rabbit anti-hu...

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Abstract

The invention discloses an application of a CGA gene as a target spot in preparation of a drug for diagnosing and treating drug-resistant solid tumors, and particularly discloses a new discovery based on high expression of the CGA in solid tumor tissues and causing drug resistance of the solid tumors, and the application of the gene or / and protein as a target spot in diagnosis and treatment of the drug-resistant solid tumors.

Description

technical field [0001] The present invention relates to solid tumor drug resistance diagnosis and prognosis evaluation technology, in particular to the new discovery based on the high expression of glycoprotein hormone alpha polypeptide (CGA, namely glycoprotein hormones, alpha polypeptide) in solid tumor tissue and lead to solid tumor drug resistance, the gene Or / and proteins are used as targets for the diagnosis and prognosis evaluation of solid tumor drug resistance. Background technique [0002] Conventional treatments for solid tumors include surgery, chemotherapy, and radiation therapy. Chemotherapy (chemotherapy) plays an important role in the treatment of solid tumors. However, most patients undergo chemotherapy drug resistance, leading to treatment failure. More importantly, tumors develop resistance to one drug (single-drug resistance) during chemotherapy, and at the same time, cross-resistance to other drugs that have not been used and have different chemical str...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886G01N33/574G01N33/74C12N5/09A01K67/027A61K45/00A61P35/00
CPCC12Q1/6886G01N33/74G01N33/574A61K45/00A61P35/00C12N5/0693A01K67/0276C12Q2600/158C12Q2600/118C12Q2600/106A01K2227/105A01K2267/0331
Inventor 赵晓迪聂勇战曹田宇卢瑗瑗樊代明
Owner FOURTH MILITARY MEDICAL UNIVERSITY