Protein biomarker for assisting in authenticating Rituximab drug-resistant ABC-DLBCL cells and application thereof

A protein marker and marker technology, applied in animal cells, tumor/cancer cells, vertebrate cells, etc., can solve the problems of poor prognosis and the inability to obtain complete remission with conventional second-line regimens.

Inactive Publication Date: 2019-11-08
CANCER INST & HOSPITAL CHINESE ACADEMY OF MEDICAL SCI
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AI Technical Summary

Problems solved by technology

Although rituximab combined with R-CHOP chemotherapy can significantly improve the outcome of DLBCL patients (the overall cure rate is about 60%), the prognosis of first-line treatment of ABC-DLBCL patients is poor (long-term progression-free survival is less than 50%), and the first-line In patients with drug resistance, more than 70% of conventional second-line regimens cannot achieve complete remission

Method used

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  • Protein biomarker for assisting in authenticating Rituximab drug-resistant ABC-DLBCL cells and application thereof
  • Protein biomarker for assisting in authenticating Rituximab drug-resistant ABC-DLBCL cells and application thereof
  • Protein biomarker for assisting in authenticating Rituximab drug-resistant ABC-DLBCL cells and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0075] Embodiment 1, the construction of drug-resistant cell line

[0076] Culture conditions: 37°C, 5% CO 2 in the cell culture incubator.

[0077] 1. The SU-DHL-2 cell line was cultured in the RPMI-1640 medium containing 10% fetal bovine serum to the logarithmic growth phase.

[0078] 2. After completing step 1, centrifuge at 1000 rpm for 5 minutes, discard the supernatant, add new RPMI-1640 medium containing 1 μg / mL Rituximab and 10% fetal bovine serum, and incubate for 24 hours.

[0079] 3. After completing step 2, centrifuge at 1000rpm for 5min, discard the supernatant, add new RPMI-1640 medium containing 10% fetal bovine serum, and culture the cells to the logarithmic growth phase.

[0080] 4. After completing step 3, centrifuge at 1000 rpm for 5 minutes, discard the supernatant, add new RPMI-1640 medium containing 1 μg / mL Rituximab and 10% fetal bovine serum, and incubate for 24 hours.

[0081] 5. After completing step 3, centrifuge at 1000rpm for 5min, discard the s...

Embodiment 2

[0106] Embodiment 2, label-free quantitative proteomics (Label-free) method

[0107] The cell samples are: cell sample A1, cell sample A2, cell sample A3, cell sample B1, cell sample B2, and cell sample B3. Cell sample A1, cell sample A2, and cell sample A3 are all SU-DHL-2-WT, representing 3 biological replicates. Cell sample B1, cell sample B2, and cell sample B3 are all SU-DHL-2-R, representing 3 biological replicates.

[0108] 1. Preparation of Protein Solution

[0109] Take cell samples, add SDT lysate, sonicate (80W, work for 10s, pause for 15s, cycle 10 times), then boil in water bath for 15min, then centrifuge at 14000g for 40min, take the supernatant, which is the protein solution.

[0110] SDT lysate: 4g / 100ml SDS, 100mM Tris-HCl, 1mM DTT, pH7.6.

[0111] Protein quantification was carried out by BCA method (BCA quantification kit, P0012, Biyuntian). Aliquot the samples and store at -80°C.

[0112] 500 μL protein solution was obtained for each cell sample. Acco...

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Abstract

The present invention discloses a protein biomarker for assisting in authenticating Rituximab drug-resistant ABC-DLBCL cells and application thereof. The application of a substance for detecting a specific biomarker to preparing a kit for detecting whether to-be-detected ABC-DLBCL cells are Rituximab drug-resistant ABC-DLBCL cells or not is protected. A biomarker A-1 is any one or a combination of8 proteins. A biomarker A-2 is any one or a combination of 45 proteins. A biomarker B-1 is any one or a combination of 5 proteins. A biomarker B-2 is any one or a combination of 77 proteins. A biomarker C is any one or a combination of 21 proteins. A biomarker D is any one or a combination of 29 proteins. According to the present invention, Rituximab drug-resistant cell strains are successfully constructed, and multiple protein biomarkers are found. The cell model and research results provide a basis for further studying Rituximab drug-resistance and overcoming drug-resistance.

Description

technical field [0001] The invention relates to a protein marker for assisting identification of Rituximab drug-resistant ABC-DLBCL cells and application thereof. Background technique [0002] Non-Hodgkin lymphoma (Non-Hodgkin lymphoma) is a common malignant tumor of lymphoid tissue, and it is one of the top 10 causes of cancer mortality. In recent years, the incidence of non-Hodgkin lymphoma in my country has also shown an obvious upward trend. Diffuse large B-cell lymphoma (DLBCL) is the most common subtype of non-Hodgkin's lymphoma (NHL), accounting for about 32.5% of newly diagnosed NHL cases each year. and other malignant tumors with great heterogeneity. DLBCL can appear in any organ and any part of the body, and the clinical manifestation is an aggressive course. According to different cell origins, DLBCL can be divided into activated B-cell-like DLBCL (ABC-DLBCL), germinal center B-cell-like DLBCL (GCB-DLBCL) and unclassified type. ABC-DLBCL has a poor prognosis and...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/68C12N5/09
CPCG01N33/56966G01N33/6848C12N5/0693C12N2501/06
Inventor 车轶群王迪刘鹏罗扬张岳沈迪曲媛王爽
Owner CANCER INST & HOSPITAL CHINESE ACADEMY OF MEDICAL SCI
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