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CN-121975738-A - Epirubicin-resistant human breast cancer cell strain, exosome and application thereof in construction of mouse chemotherapy drug-resistant breast cancer model

CN121975738ACN 121975738 ACN121975738 ACN 121975738ACN-121975738-A

Abstract

The invention belongs to the technical field of biological medicines, and particularly relates to a human breast cancer cell strain and exosome resistant to epirubicin and application thereof in construction of a mouse chemotherapy drug resistant breast cancer model, so as to solve the problem that the existing animal model is difficult to simulate the occurrence and transmission process of clinical epirubicin drug resistance. The exosome is secreted by a epirubicin-resistant human breast cancer cell line, and the epirubicin-resistant human breast cancer cell line is preserved in China Center for Type Culture Collection (CCTCC) with the preservation number of C2025338. The exosomes secreted by the cell line can reduce the accumulation of chemotherapeutic drugs in tumor tissues and up-regulate the expression of P-gp and MRP1 drug resistance proteins, thereby promoting the formation of tumor chemotherapeutic drug resistance. The mouse chemotherapy drug-resistant breast cancer model constructed by the invention is more close to the real pathological occurrence process of clinical breast cancer drug resistance, and has important basic scientific research value and application prospect for researching the chemotherapy drug-resistant mechanism, screening and reversing the chemotherapy drug-resistant drugs and exploring new targets for treatment.

Inventors

  • CHEN DONG
  • ZHANG GANLIN
  • ZUO XI
  • Nan Lingshan
  • YIN XIAOHUI

Assignees

  • 首都医科大学附属北京中医医院

Dates

Publication Date
20260505
Application Date
20260120

Claims (8)

  1. 1. The epirubicin-resistant human breast cancer cell strain is characterized by being named as an epirubicin-resistant human breast cancer cell strain MDA-MB-231/EPI, and is preserved in China center for type culture collection, the preservation number is CCTCC NO: C2025338, the preservation address is China university of Wuhan, and the preservation time is 2025, 11 and 6 days.
  2. 2. A method of establishing a strain of epirubicin-resistant human breast cancer cells according to claim 1, comprising the steps of: The stepwise concentration increasing epirubicin intermittent induction human triple negative breast cancer cell strain MDA-MB-231 cells are adopted, and stable expression epirubicin-resistant human breast cancer cell strain is obtained through multiple rounds of screening.
  3. 3. Use of exosomes in the construction of a mouse chemotherapy-resistant breast cancer model, characterized in that the exosomes are secreted by the epirubicin-resistant human breast cancer cell line of claim 1.
  4. 4. The construction method of the mouse chemotherapy drug-resistant breast cancer model is characterized by comprising the following steps of: establishing a epirubicin-resistant human breast cancer cell strain; Extracting exosomes secreted by the epirubicin-resistant human breast cancer cell line; MDA-MB-231 cells are inoculated on a mammary fat pad of an NSG mouse in situ, and a breast cancer in situ transplantation tumor model is established; when the volume of the transplanted tumor of the breast cancer in-situ transplanted tumor model reaches 80-150mm <3 >, the mouse chemotherapy drug-resistant breast cancer model is constructed by tail vein injection of the extracted exosomes.
  5. 5. The method of claim 4, wherein the epirubicin-resistant human breast cancer cell line is established by: The stepwise concentration increasing epirubicin intermittent induction human triple negative breast cancer cell strain MDA-MB-231 cells are adopted, and stable expression epirubicin-resistant human breast cancer cell strain is obtained through multiple rounds of screening.
  6. 6. Use of a mouse chemotherapy-resistant breast cancer model constructed according to the method of claim 4 or 5 for screening candidate drugs for treating breast cancer chemotherapy resistance.
  7. 7. A method of screening for a drug candidate for the treatment of breast cancer chemotherapy resistance comprising the steps of: Applying a reagent to be screened to the mouse chemotherapy drug-resistant breast cancer model constructed by the method of claim 4 or 5; and analyzing and evaluating the treatment effect of the reagent to be screened, and selecting the reagent capable of obviously improving the drug-resistant behavior of the mouse chemotherapy drug-resistant breast cancer model.
  8. 8. A candidate drug for treating breast cancer chemotherapy resistance, comprising an agent that inhibits the drug-resistant behavior of a mouse chemotherapy-resistant breast cancer model constructed by the method of claim 4 or 5.

Description

Epirubicin-resistant human breast cancer cell strain, exosome and application thereof in construction of mouse chemotherapy drug-resistant breast cancer model Technical Field The invention belongs to the technical field of biological medicines, and particularly relates to an epirubicin-resistant human breast cancer cell strain, an exosome and application thereof in construction of a mouse chemotherapy drug-resistant breast cancer model. Background Triple-negative breast cancer (TNBC) is a special and highly invasive subtype accounting for about 15% -20% of all breast cancers. Because the estrogen receptor (Estrogen receptor, ER), the progestogen receptor (Progesterone Receptor, PR) and the human EGFR 2 (Human Epidermal Growth Factor Receptor, HER 2) are negative in expression, the medicine is insensitive to endocrine treatment and HER2 targeted treatment, and besides surgical treatment, chemotherapy mainly comprising anthracyclines and other medicines is often used as a TNBC first-line treatment scheme. Epirubicin (Epirubicin, EPI) is an anthracycline commonly used in TNBC chemotherapy, however long-term chemotherapy is extremely susceptible to the development of multidrug resistance (Multidrug resistance, MDR), leading to treatment failure, tumor recurrence and distant metastasis, and is a significant challenge in clinical treatment. Chemotherapy resistance is associated with a variety of mechanisms, with overexpression of drug efflux pumps such as P-glycoprotein (P-glycoprotein, P-gp), multi-drug resistance-related protein 1 (Multidrug Resistance-Associated Protein 1, MRP 1) and the like being the classical pathway. Recent studies have shown that exosomes secreted by chemotherapy resistant cells can carry P-gp, MRP1 and specific nucleic acid molecules and deliver to sensitive cells, thereby transmitting the drug resistant phenotype inside the tumor. This exosome-mediated intercellular communication is a key way of drug resistance development in vivo. However, there is still a lack of in vivo models that effectively mimic exosome-mediated chemotherapy drug resistance delivery processes. The existing research widely adopts a subcutaneous inoculation animal model, the built tumor microenvironment has essential difference with the primary part of human breast cancer (such as breast fat pad), and the interaction of tumor cells and organ-specific microenvironment is difficult to reproduce. Therefore, the model can not simulate the natural evolution process of chemotherapy resistance in a real microenvironment, and greatly limits the exploration of related mechanisms and intervention strategies. Therefore, the animal model which can simulate the mammary gland specific microenvironment and exosome mediated drug resistance transmission simultaneously is constructed, and is important for truly reducing the occurrence and development mechanism of clinical chemotherapy drug resistance. Disclosure of Invention The invention aims to overcome the defects of the prior art and provide a epirubicin-resistant human breast cancer cell strain, an exosome and application thereof in constructing a mouse chemotherapy drug-resistant breast cancer model. The technical idea of the invention is as follows: By in situ seeding MDA-MB-231 cells on mouse mammary gland fat pad, the microenvironment comprises adipocytes, fibroblasts, immune cells and specific extracellular matrix, and the primary planting and growing conditions of human breast cancer can be highly simulated. On the basis, the exosomes derived from the chemotherapy drug-resistant cells are used for intervention, so that the in-situ tumor can be driven to generate acquired drug resistance. The model constructed by the combined strategy not only keeps the influence of the in-situ microenvironment on tumor evolution, but also can more completely simulate the natural pathological process of the clinical TNBC for generating acquired drug resistance through intercellular communication under the microenvironment of a specific organ due to the introduction of an exosome which mediates intercellular communication, thereby providing a highly clinically relevant reliable research platform for deeply researching the drug resistance mechanism of the TNBC and developing a new method for reversing the drug resistance. The first aspect of the invention provides a human breast cancer cell strain resistant to epirubicin, which is named as a human breast cancer cell strain MDA-MB-231/EPI Homo sapiens resistant to epirubicin, and is preserved in China center for type culture Collection with the preservation number of CCTCC NO: C2025338, the preservation address of China university of Wuhan and Wuhan, and the preservation time of 2025 years, 11 months and 6 days. The second aspect of the invention provides a method for establishing the epirubicin-resistant human breast cancer cell strain, which comprises the following steps: The stepwise concentration increasing epirubicin intermittent inducti