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Markers of drug resistance of cancer cells, combination of preparations for reversing drug resistance of cancer cells and their applications

A cancer cell, drug-resistant technology, applied in the field of oncology, can solve problems such as impact

Active Publication Date: 2022-03-18
TONGJI HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Although PARP inhibitors are gradually used in the treatment of ovarian cancer, the ensuing drug resistance is also the main obstacle to further improve the prognosis of patients. The drug resistance of PARP inhibitors may be attributed to the secondary inheritance of tumor cells themselves. event or epigenetic event

Method used

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  • Markers of drug resistance of cancer cells, combination of preparations for reversing drug resistance of cancer cells and their applications
  • Markers of drug resistance of cancer cells, combination of preparations for reversing drug resistance of cancer cells and their applications
  • Markers of drug resistance of cancer cells, combination of preparations for reversing drug resistance of cancer cells and their applications

Examples

Experimental program
Comparison scheme
Effect test

experiment example 1

[0050]Experimental example 1 Ovarian cancer cell line culture

[0051] Ovarian cancer cell lines and medium: OV90, OVCAR3, A2780, OVCAR8 and SKOV3 (all purchased from ATCC, USA) were cultured in McCoy's 5A medium (Gibco, USA), and HCC1937 and SW626 were cultured in L15 (Gibco, USA) MRC5 was cultured in MEM (Gibco, USA) medium.

[0052] All of the above growth media were supplemented with 1% penicillin / streptomycin mix mixed with 10% fetal calf serum. The cells were in an adherent growth state and were cultured at 37°C with 5% CO 2 in the incubator.

experiment example 2

[0053] Experimental Example 2 Isolation and Purification of Human Ovarian Cancer-Associated Fibroblasts (CAFs)

[0054] Take 10g of fresh ovarian cancer tissue (collected from the Department of Obstetrics and Gynecology, Tongji Hospital Affiliated to Tongji Medical College, Huazhong University of Science and Technology), rinse with normal saline for 3 times, and cut the tissue to 1mm under sterile conditions on the cell workbench 3 size. Transfer the shredded tissue pieces to a 50ml sterile centrifuge tube, add 20ml tissue digestion solution, and digest in a shaker at 37°C for 2-4h. The tissue digestion solution was prepared from serum-free DMEM / F12 (Thermo Fisher) medium, containing type I collagenase (Sigma Company) with a final concentration of 1 μg / ml and type I DNAase (Roche Company) with a final concentration of 1 μg / ml , 1% penicillin / streptomycin mixture. Stop the digestion when the tissue block shrinks significantly and the digestive juice becomes turbid, and centri...

experiment example 3

[0056] Experimental Example 3 Drug Sensitivity Determination

[0057] The CAFs cells and MRC5-CAFs obtained in Experimental Example 2 were respectively digested with trypsin to prepare single-cell suspensions, and the cells were divided into 1×10 cells per well. 4 inoculated into a sterile 96-well plate.

[0058] After 24 hours, the medium solution containing olaparib or niraparib was used to replace the well plate. After 48 hours, the cells were rinsed with PBS, and 100 μl of serum-free medium (Gibco, USA) containing 10% CCK-8 was added to each well. The 96-well plate was wrapped with light-proof paper to avoid light, and after being incubated in an incubator for 2 h, the absorbance of each well at a wavelength of 450 nm was measured with a microplate reader (Bio-Rad Laboratories). The calculation of cell activity is as follows: cell activity (%)=(drug group-blank well) OD450 / (no drug group-blank well) OD450×100%. Three secondary wells were set up for each treatment. The ...

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Abstract

The invention relates to a marker of drug resistance of cancer cells, a combination of preparations for reversing drug resistance of cancer cells and applications thereof. The markers include CCL5 chemokines of cancer-associated fibroblasts and genes encoded thereof, and the expression level of the CCL5 chemokines is positively correlated with drug resistance of cancer cells. The preparation combination includes a PARP inhibitor for inhibiting the replication of cancer cells and a reversal agent for regulating the drug resistance of cancer cells caused by the PARP. The reversal agent includes at least one of regulators for downregulating the expression of CCL5 in cancer-associated fibroblasts, CCL5 neutralizing antibodies and NF-κB signal inhibitors. Down-regulation of CCL5 expression-related regulators, CCL5 neutralizing antibodies and / or targeting NF-κB inhibitors can reverse PARP inhibitor-induced CAFs activation, inhibit PARP inhibitor-induced cancer cell resistance, and improve PARP inhibitor killing effect on cancer cells.

Description

technical field [0001] The invention relates to the technical field of oncology, in particular to a marker of drug resistance of cancer cells, a combination of preparations for reversing drug resistance of cancer cells and applications thereof. Background technique [0002] Polyadenosine diphosphate-ribose polymerase (PARP) inhibitor is a medical drug that can affect the way cancer cells replicate themselves. It can work together with other breast cancer drugs to fight cancer. This drug can also treat ovarian cancer, Prostate cancer, pancreatic cancer and other hereditary cancers that share the same "rogue gene"; it can also be used as a new type of molecular targeted drug to improve the prognosis of ovarian cancer patients and apply to clinical treatment. [0003] PARP inhibitors mainly act on tumor cells with homologous recombination repair gene defects, resulting in DNA damage that cannot be repaired, thereby triggering the accumulation of DNA damage and ultimately cell d...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886G01N33/574G01N33/68A61K45/06A61P35/00
CPCC12Q1/6886G01N33/57449G01N33/57415G01N33/57434G01N33/57438G01N33/57488G01N33/6863A61K45/06A61P35/00C12Q2600/158C12Q2600/106G01N2333/523
Inventor 高庆蕾马丁杨宗元李晓婷徐森
Owner TONGJI HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI TECH
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