Unlock instant, AI-driven research and patent intelligence for your innovation.

New application of sialyltransferase ST3GAL4 as EGFR-TKI drug resistance marker

A technology of EGFR-TKI and sialyltransferase, applied in the field of biomedicine, can solve problems such as limited clinical application, acquired drug resistance, and complex drug resistance mechanism

Pending Publication Date: 2022-04-15
中国人民解放军陆军特色医学中心
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, EGFR-TKIs such as osimertinib, gefitinib, and afatinib also face the problem of acquired drug resistance, and the drug resistance mechanism is complex, which severely limits their clinical application

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • New application of sialyltransferase ST3GAL4 as EGFR-TKI drug resistance marker
  • New application of sialyltransferase ST3GAL4 as EGFR-TKI drug resistance marker
  • New application of sialyltransferase ST3GAL4 as EGFR-TKI drug resistance marker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Embodiment 1, immunoblotting experiment (WB) detects the expression of drug-resistant cell sialyltransferase ST3GAL4

[0046] 1. Experimental materials:

[0047] RPMI 1640 medium, fetal bovine serum, penicillin-streptomycin solution, EDTA trypsin, cell culture flask, cell culture dish, tissue protein lysate, BCA protein concentration assay kit, 30% Acr-Bis (29:1) , Tris-Hcl (PH8.8 and PH6.8), 10% SDS, 10% ammonium persulfate, TEMED, 5× protein loading buffer, ST3GAL4 antibody, GAPDH antibody (internal reference antibody), Western primary antibody diluent, Protein Marker, skimmed milk powder, goat anti-rabbit secondary antibody, TRIS, SDS, Glycine, sodium chloride, Tween-20.

[0048] Cell culture medium: RPMI 1640 medium + 10% fetal bovine serum + 0.1% penicillin-streptomycin solution

[0049] 10× electrophoresis solution: SDS 10g, Glycine 188g, Tris 30.2g were dissolved in 1L distilled water. Dilute with distilled water to 1× electrophoresis solution before use.

[...

Embodiment 2

[0071] Test results such as figure 1 As shown, compared with osimertinib-sensitive cell PC-9GR, drug-resistant cell lines PC-9GROR1, PC-9GROR2, PC-9GROR3 and H1975OR2, H1975OR3 sialyltransferase ST3GAL4 expression enhanced. Example 2. Enzyme-linked immunosorbent assay (ELISA) detects the concentration change of sialyltransferase ST3GAL4 in the patient's serum samples before and after osimertinib resistance.

[0072] 1. Experimental materials

[0073] Human ST3GAL4 enzyme-linked immunoassay kit, patient's serum.

[0074] 2. Experimental steps

[0075] (1) Dilution of standard

[0076] This kit provides an original standard, which is diluted in a small test tube according to the following table.

[0077] 240pg / ml Standard No. 5 Add 150μl standard diluent to 150μl original standard 120pg / ml Standard No. 4 Add 150μl of Standard No. 5 to 150μl of Standard Diluent 60pg / ml Standard No. 3 Add 150μl of Standard No. 4 to 150μl of Standard Diluent ...

Embodiment 3

[0103] 1. Obtain PC-9GR stable cell line and H1975 stable cell line overexpressing Human ST3GAL4 gene

[0104] First, PC-9GR and H1975 cells overexpressed ST3GAL4, and the following steps were followed: using lentivirus as a tool, the exogenous gene Human ST3GAL4 (C-terminal GFP tag) was integrated into PC-9GR cell line and H1975 cell line respectively, and the obtained PC-9GR stable cell line and H1975 stable cell line expressing Human ST3GAL4 gene. (Human ST3GAL4 gene information GenBank: BC010645.1).

[0105] (1) Vector construction and extraction

[0106] Synthesize the target gene ST3GAL4 (fused with C-terminal GFP tag), and insert the synthetic sequence into the lentiviral transfer vector (vector name pLV-ST3GAL4, eukaryotic resistance is puromycin, prokaryotic resistance is Ampicillin). Pick multiple clones for sequencing identification, select the correct activated strain, transfer to LB medium, and shake the bacteria overnight at 37°C. After overnight culture, extr...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses an application of sialyltransferase ST3GAL4 serving as a marker for assisting in evaluating EGFR-TKI (epigallocatechin gallate receptor-TKI) drug resistance, an application of the ST3GAL4 in preparing a reagent serving as the marker for evaluating the EGFR-TKI drug resistance, and an application of the ST3GAL4 in preparing a kit for assisting in evaluating the EGFR-TKI drug resistance. The invention further discloses an EGFR-TKI drug resistance analysis kit, and the EGFR-TKI drug resistance analysis kit comprises the ST3GAL4 expression quantity detection reagent. Experiments prove that ST3GAL4 is closely related to EGFR-TKI drug resistance, ST3GAL4 can be used as an EGFR-TKI drug resistance marker, ST3GAL4 detection has a mature technology at present, for example, an ELISA kit is used for detecting a patient blood sample, IHC is used for detecting a patient tissue wax block sample, and by detecting ST3GAL4, whether a patient is resistant to EGFR-TKI can be indicated, and a treatment strategy can be guided and adjusted in time.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a new application of sialyltransferase ST3GAL4 as an EGFR-TKI drug resistance marker and an EGFR-TKI drug resistance analysis kit. Background technique [0002] Lung cancer is divided into non-small cell lung cancer and small cell lung cancer, of which non-small cell lung cancer accounts for more than 80%. [0003] Osimertinib (OSI), as the third epidermal growth factor receptor-tyrosine kinase inhibitor (EGFR-TKI), is currently widely used in advanced non-small cell lung cancer with EGFR sensitive mutations The first-line and second-line treatment of patients have remarkable curative effect, and it is the representative drug of the third-generation EGFR-TKI. Gefitinib (Gefitinib, Yiruike, Iressa) is an oral epidermal growth factor receptor tyrosine kinase (EGFR-TK) inhibitor (a small molecule compound), which is the first generation of EGFR-TKI represents drugs. The first...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): G01N33/573G01N33/58
Inventor 何勇韩睿
Owner 中国人民解放军陆军特色医学中心