Human mammary gland malignant leafy tumor cell line SHSY-MPT-06 as well as preparation method and application thereof

Abstract

The invention relates to a human mammary gland malignant leaky tumor cell line SYSH-MPT-06 as well as a preparation method and application thereof, and belongs to the technical field of medicine. The human mammary gland malignant leafy tumor cell line is established from a Chinese human source, is short in establishment time and stable in biological heredity, can form an in-situ tumor under a mammary gland fat pad of a nude mouse, can form a lung metastatic tumor through a caudal vein of the mouse, has tumor formation capability and metastasis capability, and can be used for preparing a tumor cell. As a research model, the human mammary gland malignant leaky tumor cell line is greatly helpful for knowing pathogenesis, signal channels and tumor occurrence, development or metastasis of patients with mammary gland leaky tumors, and can also be used for screening and evaluating medicines for resisting mammary gland leaky tumors and preparing tumor diagnosis products. The method has high scientific research and production application values, and is expected to produce good scientific research, economic and social benefits.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and particularly to a human malignant phyllodes tumor cell line SHSY-MPT-06, a preparation method thereof, and an application thereof. Background Art

[0002] Breast phyllodes tumor (PT) is a fibroepithelial tumor with a phyllodes structure, composed of breast stromal fibroconnective tissue and epithelial tissue, accounting for less than 1% of all breast tumors. According to the clinicopathological characteristics of phyllodes tumors, the World Health Organization (WHO) classifies them into three categories: benign, borderline, and malignant phyllodes tumors. Among them, compared with benign and borderline phyllodes tumors, malignant phyllodes tumors are characterized by rapid growth and strong invasiveness. Pathological examinations show that malignant phyllodes tumors have rich blood supply, infiltrative growth, obvious nuclear atypia, active mitosis, and excessive stromal growth. In addition, malignant phyllodes tumors have a high recurrence and metastasis rate and are insensitive to chemotherapy, radiotherapy, endocrine therapy, targeted therapy, and immunotherapy. Surgical treatment remains the main treatment method at present. Generally speaking, the prognosis of malignant phyllodes tumors is poor, and the overall survival rate is low. There is an urgent need to seek more effective drugs through in-depth scientific research to further improve the quality of life of patients with malignant phyllodes tumors.

[0003] Human breast phyllodes tumor cell lines are commonly used tools in the study of phyllodes tumors. Human breast phyllodes tumor cell lines retain the biological behaviors and characteristics of breast phyllodes tumors, can be continuously passaged in vitro, and the responses of cells to various drug treatments can be monitored in real time by cell biological means. At the same time, very few malignant phyllodes tumor cell lines can form orthotopic xenografts in experimental animals, and currently, there is a lack of human malignant phyllodes tumor cell lines that can form lung metastasis models in experimental animals by tail vein injection. These cell lines can be used for the mechanism research of the occurrence and malignant progression of breast malignant phyllodes tumors and the systematic evaluation of treatments.

[0004] Different from breast cancer with a high incidence rate, there are few studies on the occurrence, malignant progression, and their mechanisms of breast phyllodes tumors. And due to the large heterogeneity of phyllodes tumors, phyllodes tumor cell lines derived from different patients usually show different tumor biological characteristics. This heterogeneity requires researchers to try to use multiple phyllodes tumor cell lines during the research process to ensure that the research results have a wide range of applicability. At the same time, there is currently no human breast phyllodes tumor cell line that can simultaneously carry out in vitro passage, orthotopic breast xenografts, and lung metastasis models. There is an urgent need to establish a new breast phyllodes tumor cell line that can establish multiple in vivo and in vitro models to ensure that researchers can cover more types of breast malignant phyllodes tumors. Summary of the Invention

[0005] The object of the present invention is to overcome the deficiencies of the prior art and provide a human malignant phyllodes tumor cell line SHSY-MPT-06, its preparation method and applications.

[0006] To achieve the above object, the technical solution adopted by the present invention is as follows:

[0007] In the first aspect, the present invention provides a human malignant phyllodes tumor cell line, named human malignant phyllodes tumor cell line SHSY-MPT-06, which is deposited in the China Center for Type Culture Collection (CCTCC), the deposit address is: Wuhan University, Wuhan, China, and the deposit number is CCTCC NO: C202584.

[0008] The present invention obtains a human malignant phyllodes tumor cell line SYSH-MPT-06 from fresh surgical samples of malignant phyllodes tumor tissues through steps including construction of a patient-derived tumor xenograft (PDX) model, passage of the PDX model, primary cell culture and cell passage culture. The cells of this cell line are polygonal in shape and show the morphological characteristics of fibroepithelial cells; and it is observed that the cells grow malignantly but still maintain contact inhibition. Orthotopic tumors can be formed under the mammary fat pad of nude mice, and lung metastatic tumors can be formed through the tail vein of mice. This cell has the ability to form tumors and the ability to metastasize. Therefore, the human malignant phyllodes tumor cell line SYSH-MPT-06 of the present invention is deposited, and the deposit address of this cell line is: Wuhan University, Wuhan, China, and the deposit number is CCTCC NO: C202584.

[0009] In the second aspect, the present invention provides the application of the above-mentioned human malignant phyllodes tumor cell line in establishing an animal or cell model of human malignant phyllodes tumor.

[0010] In the third aspect, the present invention provides the application of the human malignant phyllodes tumor cell line in in vitro research on the pathogenic mechanism, related signaling pathways, and tumorigenesis, development or metastasis mechanism of human malignant phyllodes tumor.

[0011] In the fourth aspect, the present invention provides the application of the human malignant phyllodes tumor cell line in screening diagnostic products for human malignant phyllodes tumor.

[0012] In the fifth aspect, the present invention provides the application of the human malignant phyllodes tumor cell line in screening and detecting bioengineering products related to human malignant phyllodes tumor.

[0013] Sixth aspect, the present invention provides the application of a human malignant phyllodes tumor cell line in screening drug targets for human malignant phyllodes tumors.

[0014] Seventh aspect, the present invention provides the application of a human malignant phyllodes tumor cell line in screening or evaluating anti-human malignant phyllodes tumor drug reagents.

[0015] Eighth aspect, the present invention provides a cell model, and the cell model is the cell line of the human malignant phyllodes tumor cell line described above.

[0016] Ninth aspect, the present invention provides a method for constructing the human malignant phyllodes tumor cell line described above, which is characterized by including the following steps:

[0017] (1) Inoculate a human malignant phyllodes tumor tissue mass under the mammary fat pad of a mouse;

[0018] (2) After culturing the mouse, take out the tumor tissue;

[0019] (3) Then inoculate the tumor tissue mass in step (2) into the mammary fat pad of the mouse for passage of the PDX model;

[0020] (4) After the PDX model has been passaged five times, take out the tumor tissue for primary cell culture;

[0021] (5) When the cells in step (4) grow to 80% confluence, perform cell passage culture;

[0022] (6) After passage to the 50th generation in step (5), the human malignant phyllodes tumor cell line described above is obtained.

[0023] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0024] The human malignant phyllodes tumor cell SYSH-MPT-06 of the present invention can be passaged in vitro under the conditions of conventional cell culture, has stable in vitro characteristics, and conforms to the clinical tumor biological characteristics. The human malignant phyllodes tumor cell line has tumorigenicity and can be used to prepare an orthotopic transplantation tumor model of malignant phyllodes tumor of the breast and a lung metastasis model of malignant phyllodes tumor of the breast, both of which can be used for the mechanism research of the occurrence and development of phyllodes tumor of the breast and the in vivo evaluation of drug effects. The malignant phyllodes tumor cell line of the breast forms breast in-situ tumors and lung metastases quickly, which is beneficial to the development of scientific research. In addition, the malignant phyllodes tumor cell line of the breast can be used for the in vitro evaluation of the drug effects of malignant phyllodes tumors of the breast. Generally speaking, the human malignant phyllodes tumor cell line can be used to guide clinical medication, is beneficial to the discovery of new anti-malignant phyllodes tumor drugs, and has good scientific research value and economic benefits. Brief Description of the Drawings

[0025] Figure 1 It is the cell morphology diagram of cell SYSH-MPT-06 under light microscope;

[0026] Figure 2 It is the cell growth curve diagram of cells SYSH-MPT-06, LJ-0429, and HJP-0320;

[0027] Figure 3 It is the cell viability-drug dose curve diagram of cells SYSH-MPT-06, LJ-0429, HJP-0320 (left), and docetaxel (right);

[0028] Figure 4 It is the schematic diagram of the results of the Transwell migration experiment (upper) and invasion experiment (lower) of cells SYSH-MPT-06, LJ-0429, and HJP-0320;

[0029] Figure 5 It is the schematic diagram of the results of the in-situ tumorigenesis experiment of cells SYSH-MPT-06, LJ-0429, and HJP-0320 in nude mice;

[0030] Figure 6 It is the growth curve diagram of the in-situ transplanted tumor of cell SYSH-MPT-06;

[0031] Figure 7 It is the body weight curve diagram of nude mice with tail vein metastatic tumors constructed with cells SYSH-MPT-06, LJ-0429, and HJP-0320;

[0032] Figure 8 It is the typical schematic diagram of the in-situ transplanted tumor of cell SYSH-MPT-06;

[0033] Figure 9 It is the schematic diagram of the lungs of nude mice with lung metastasis models constructed with cells SYSH-MPT-06, LJ-0429, and HJP-0320;

[0034] Figure 10 It is the schematic diagram of the HE staining results of the lung metastasis lesions in the lungs of mice with lung metastasis models constructed with cells SYSH-MPT-06, LJ-0429, and HJP-0320. Detailed implementation manners

[0035] To better illustrate the purpose, technical solution, and advantages of the present invention, the present invention will be further described below in conjunction with specific embodiments.

[0036] The present invention obtains a human malignant phyllodes tumor cell line SYSH-MPT-06 from a fresh surgical sample of human malignant phyllodes tumor tissue in China through steps of PDX model construction, PDX model passage, primary cell culture, and cell passage culture. The cells of this cell line are polygonal in shape and exhibit the morphological characteristics of fibroepithelial cells. It is observed that the cells grow malignantly but still maintain contact inhibition. Orthotopic tumors can be formed under the mammary fat pads of nude mice, and lung metastatic tumors can be formed through the mouse tail vein. This cell has the ability to form tumors and the ability to metastasize. Therefore, the present invention deposits the human malignant phyllodes tumor cell line SYSH-MPT-06. The cell line is deposited at the China Center for Type Culture Collection (CCTCC), with the deposit address: Wuhan University, Wuhan, China, and the deposit number: CCTCC NO: C202584; the deposit date: March 6, 2025.

[0037] Example 1 Preparation of the human malignant phyllodes tumor cell line SYSH-MPT-06

[0038] Construction of a patient-derived mouse xenograft tumor model (PDX, Patient-Derived Tumor Xenograft): Cut a fresh surgical sample of malignant phyllodes tumor tissue with a size of 1 cm × 1 cm × 1 cm obtained in a laminar flow operating room (pathological result: In the breast tissue (right breast), tumor cells grow diffusely in sheets. The tumor cells are spindle-shaped or oval, tumor giant cells can be seen, mitotic figures are easily seen, accompanied by large areas of necrosis. Osteoid and chondroid matrix can be seen in some areas, and a small amount of benign glandular ducts are interspersed in focal areas. The tumor invades the skin epidermis with ulcer formation, and infiltrating striated muscle tissue is seen. Tumors can be seen at the basal cauterized margin, no definite intravascular tumor thrombus is seen, and no tumor is seen in the nipple. Combining the immunohistochemical results, it is in line with malignant phyllodes tumor with heterologous differentiation.) and store it in 20 ml of sterile DMEM culture medium at 4°C, and transfer it to the barrier system animal room within 24 hours for the following aseptic operations. First, wash the malignant phyllodes tumor tissue twice with sterile PBS at 4°C, then cut it into tissue blocks with a size of 1 mm × 1 mm × 1 mm, inoculate the tumor tissue blocks under the second pair of mammary fat pads of anesthetized four-week-old female NOD / SCID mice, suture the incision, and wait for the mice to wake up and observe for a period of time.

[0039] Passage of the patient-derived mouse xenograft tumor model (PDX): When the tumor volume in the mouse reaches 1000 mm^3, euthanize the mouse in the barrier by carbon dioxide asphyxiation, take out the tumor tissue, wash it three times with sterile PBS at 4°C, then cut it into tissue blocks with a size of 1 mm × 1 mm × 1 mm, and re-inoculate the tissue blocks under the second pair of mammary fat pads of anesthetized four-week-old female NOD / SCID mice, suture the incision, and perform the passage of the PDX model.

[0040] Primary cell culture: After the PDX model has been passaged five times, when the tumor volume of the fifth passage reaches 1000 mm^3, the mice are euthanized by carbon dioxide, the tumor tissues are removed, and the tumor tissue samples are immediately stored in 20 ml of sterile DMEM culture medium at 4 °C and transported on ice to the biosafety cabinet within 12 hours for the following aseptic operations. First, the mammary gland malignant phyllodes tumor tissue is mechanically minced as finely as possible, and then digested with 20 ml of tissue digestion solution (DMEM F12 culture medium containing 1 mg / ml of type III collagenase, 100 U / ml of penicillin, and 100 μg / ml of streptomycin sulfate) under shaking conditions at 37 °C for 2 hours, wrapped with tin foil to avoid light. After filtering the digested cell suspension through a 70 μm filter to obtain a single-cell suspension, it is centrifuged at 2000 rpm for 20 min, the supernatant is discarded, and the cell pellet is obtained. After washing the cells twice with 0.9% physiological saline, they are resuspended in 1 ml of complete medium (DMEM F12 culture medium containing 16% fetal bovine serum, 100 U / ml of penicillin, and 100 μg / ml of streptomycin sulfate), seeded in a 10 cm dish, 8 ml of complete medium is added, and it is statically cultured in an incubator at 37 °C and 5% (V / V) CO 2 for cell culture. After 24 hours, the medium is changed and the non-adherent cells are washed away. Thereafter, the above-mentioned culture medium is changed every 2 days.

[0041] Cell subculture: When the cells grow to 80% confluence, cell subculture is carried out in the biosafety cabinet. The specific steps are as follows: Discard the old medium, wash the cells with sterile PBS with a pH value of 7.4 and discard, add 1.5 ml of trypsin solution to digest until the cells are suspended, add 1.5 ml of culture medium containing 10% serum to terminate digestion, and collect all the cells into a 15 ml centrifuge tube. After centrifuging at 1000 rpm for 5 minutes, the supernatant is discarded. Then the cells are resuspended with fresh culture medium and passaged and inoculated in a new 10 cm dish at a ratio of 1:3. For the first 15 passages, DMEM F12 culture medium containing 16% fetal bovine serum, 100 U / ml of penicillin, and 100 μg / ml of streptomycin sulfate is used. After passage to the 15th generation, DMEM F12 culture medium containing 10% fetal bovine serum, 100 U / ml of penicillin, and 100 μg / ml of streptomycin sulfate can be used for culture.

[0042] It was observed after culture that the mammary gland malignant phyllodes tumor cells showed a fibroepithelial-like structure, with uniform cell morphology, contact inhibition, and a stable cell growth rate. The cells were named human mammary gland malignant phyllodes tumor cells SYSH-MPT-06.

[0043] Example 2 Biological characteristics and applications of human mammary gland malignant phyllodes tumor cells

[0044] Morphological observation: Place a 10-cm dish containing SYSH-MPT-06 cells under an inverted microscope for observation and photography. As Figure 1 shown, the cells are polygonal in shape, showing the morphological characteristics of fibroepithelial cells; it was also observed that the cells grew malignantly but still maintained contact inhibition.

[0045] Determination of cell doubling time: Seed the cells in a 96-well plate at a density of 3000 cells / well. Measure the cell viability by the CCK-8 method at 0, 24, 48, 72, and 96 hours. According to the absorbance value curve ( Figure 2 ), it can be seen from the proliferation curve that under the same conditions, the proliferation rate of SYSH-MPT-06 is between that of the commonly used breast malignant phyllodes tumor cell lines LJ-0429 and HJP-0320.

[0046] Determination of the half-lethal dose IC50 of doxorubicin and docetaxel: Seed the cells in a 96-well plate at a density of 5000 cells / well. After 24 hours, add doxorubicin and docetaxel at different concentrations. After 72 hours of drug treatment, measure the cell viability by the CCK-8 method, and calculate the half-lethal dose of the drug based on the drug concentration and cell viability. The results are as Figure 3 shown. The doxorubicin IC50 of SYSH-MPT-06 is 495.8 nM, higher than that of the commercial breast malignant phyllodes tumor cell lines LJ-0429 (259.5 nM) and HJP-0320 (87.76 nM); the docetaxel IC50 of SYSH-MPT-06 is 200.4 nM, higher than that of the commercial breast malignant phyllodes tumor cell lines LJ-0429 (0.7344 nM) and HJP-0320 (3.646 nM).

[0047] Evaluation of in vitro migration and invasion abilities: To evaluate the migration ability of the cells, place a cell transwell chamber with an 8-μm pore size in a 24-well plate. Add 100 μl of serum-free culture medium containing 10,000 cells to the upper chamber (the upper space of the chamber), and add 800 μl of culture medium containing 20% fetal bovine serum to the lower chamber (the external space of the chamber). After static culture in a cell incubator for 24 hours, take out the chamber, wipe off the cells on the upper side of the bottom membrane of the chamber with a cotton swab, fix the cells on the lower side of the bottom membrane of the chamber with 4% paraformaldehyde solution and stain with crystal violet, and observe the number of cells migrated to the lower side of the bottom membrane under an optical microscope. The evaluation method for the invasion ability is similar, except that for the invasion experiment, 50 μl of 1:8 diluted Matrigel needs to be coated on the cell transwell chamber in advance, and the subsequent operation method is exactly the same as that of the migration experiment. The experimental results ( Figure 4 ) show that the migration and invasion abilities of SYSH-MPT-06 are stronger than those of the commercial human breast malignant phyllodes tumor cell lines LJ-0429 and HJP-0320.

[0048] Evaluation of in vivo orthotopic tumorigenicity: For the orthotopic tumor formation experiment, female BALB / c-Nude nude mice aged 4 - 6 weeks were used. The cells of three breast malignant phyllodes tumor cell lines, SYSH-MPT-06, LJ-0429, and HJP-0320, were resuspended in PBS. After adjusting the cell concentration to 2×10^7 cells / ml, they were mixed with Matrigel in equal volume to obtain a cell / Matrigel mixture with a cell concentration of 1×10^7 cells / ml. 0.15 ml of the cell / Matrigel mixture was injected under the second pair of mammary fat pads on the left side of the nude mice. The injection site was observed every 4 days. After 12 days of inoculation, it could be observed that tumors gradually formed under the armpits of the nude mice inoculated with SYSH-MPT-06, and prominent masses could be observed subcutaneously in the nude mice with the naked eye, while no tumors formed in the nude mice inoculated with LJ-0429 and HJP-0320( Figure 5 ). The long axis diameter and short axis diameter (perpendicular to each other) of the subcutaneous tumors were measured with a vernier caliper, and the tumor volume was calculated according to "tumor volume = 1 / 2 × long axis diameter × short axis diameter × short axis diameter". On the 28th day after inoculation, the average tumor volume in the SYSH-MPT-06 group reached 1000 cubic millimeters( Figure 6 ). The experimental results showed that compared with the commercial malignant phyllodes tumor cell lines LJ-0429 and HJP-0320, SYSH-MPT-06 had the ability to rapidly form orthotopic tumors in the mammary fat pads of mice( Figure 8 ) ， However, the malignant phyllodes tumor cell lines LJ-0429 and HJP-0320 could not form orthotopic tumors in the mammary fat pads of mice.

[0049] Evaluation of lung metastasis ability: For the lung metastasis model via the tail vein, female nude mice aged 4 - 6 weeks were used. The cells of three malignant phyllodes tumor cell lines, SYSH-MPT-06, LJ-0429, and HJP-0320, were respectively resuspended in PBS. After adjusting the cell concentration to 5×10^6 cells / ml, 200 μl of the cell suspension was injected into the mice through the tail vein of the mice using a micro syringe with an insulin needle. After the injection, the needle was withdrawn after pressing for ten seconds, and the mice were put back into the cage. The body weight of the mice was weighed and recorded every four days. The body weight curves of the mice in each group were as Figure 7 shown. After 16 days of inoculation, it could be observed that the growth rate of the body weight of the mice in the SYSH-MPT-06 group slowed down significantly. On the 44th day after inoculation, the mice were sacrificed using an euthanasia instrument, and the lungs were isolated. The lung pictures of the mice in each group were photographed and recorded. The experimental results were as Figure 9As shown. After fixation with 4% paraformaldehyde, paraffin-embedded sections of the lungs of mice in each group were prepared and HE staining was performed. The results showed that metastatic foci were formed only in the lungs of nude mice injected with SYSH-MPT-06 cells via the tail vein, while no metastatic foci could be formed in the lungs of nude mice after injection of cell lines LJ-0429 and HJP-0320( Figure 10 ).

[0050] Short tandem repeats (STR) identification: STR (Short Tandem Repeat) gene loci are composed of short tandem repeat sequences with a length of 3 to 7 base pairs. These repeat sequences are widely present in the human genome and can be used as highly polymorphic markers, known as the DNA fingerprint of cells. The identification process is as follows: Genomic DNA of SYSH-MPT-06 was extracted, the STR sequence was amplified by polymerase chain reaction, the amplified product was separated by capillary electrophoresis, the copy number of the STR sequence was identified by fluorescence detection, and finally compared with the cell STR database to deduce the name of the cell line to which it belongs or the possible cross-contaminated cell line. STR identification was performed on human mammary malignant phyllodes tumor cells SYSH-MPT-06 and its primary cells (Guangzhou Aiji Biotechnology Co., Ltd.). The results are shown in Table 1. The copy number of the SYSH-MPT-06 cell line of the present invention at 21 STR loci completely matches that of the primary cells, indicating that the SYSH-MPT-06 cell line of the present invention is homologous to the primary cells and has not been contaminated by other cells.

[0051] Table 1 STR identification results of HCCP-ZJ

[0052]

[0053]

[0054] In summary, the SYSH-MPT-06 cells cultured in DMEM / F12 medium containing 10% fetal bovine serum in the present invention achieved long-term growth in vitro and stable passage. Observed under a microscope, SYSH-MPT-06 has a typical fibroepithelial cell-like morphology. The results of the CCK8 experiment showed that the proliferation rate of SYSH-MPT-06 was between LJ-0429 and HJP-0320, the half-lethal dose (IC50) of doxorubicin was 495.8 nM, and the half-lethal dose of docetaxel was 200.4 nM. The perforation experiment and invasion experiment showed that SYSH-MPT-06 exhibited strong migration and invasion abilities in vitro. A certain amount of SYSH-MPT-06 cells could form orthotopic tumors under the mammary fat pad of nude mice and lung metastatic tumors through the tail vein of mice, indicating that the cells have tumorigenic ability and metastatic ability.

[0055] However, compared with the cell line SYSH-MPT-06 of the present invention, commercialized malignant phyllodes tumor cell lines such as LJ-0429 and HJP-0320 do not have the ability to form tumors in vivo; at the same time, they are also unable to form lung metastases in immunodeficient mice. Accordingly, compared with previously reported human breast malignant phyllodes tumor cell lines, SYSH-MPT-06 significantly fills the gap in the use of malignant phyllodes tumor cell lines for in vivo modeling, and can provide great significance for the in-depth study of breast malignant phyllodes tumor and the improvement of its diagnosis and treatment prognosis.

[0056] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention rather than to limit the protection scope of the present invention. Although the present invention has been described in detail with reference to the preferred embodiments, those of ordinary skill in the art should understand that the technical solutions of the present invention can be modified or equivalently replaced without departing from the essence and scope of the technical solutions of the present invention.

Claims

1. A human breast malignant phyllodes tumor cell line, characterized in that The human breast malignant phyllodes tumor cell line was named SHSY-MPT-06, and the cell line was deposited in China Center for Type Culture Collection, with the deposit address being Wuhan University, Wuhan, China, and the deposit number being CCTCC NO: C202584.

2. Use of the human breast malignant phyllodes tumor cell line according to claim 1 in establishing an animal or cell model of human breast malignant phyllodes tumor.

3. Use of the human breast malignant phyllodes tumor cell line according to claim 1 in in vitro studies of the pathogenic mechanism, related signaling pathways, and tumor occurrence, development, or metastasis mechanism of human breast malignant phyllodes tumor.

4. Use of the human breast malignant phyllodes tumor cell line according to claim 1 in screening diagnostic products for human breast malignant phyllodes tumor.

5. Use of the human breast malignant phyllodes tumor cell line according to claim 1 in screening and detecting bioengineering products related to human breast malignant phyllodes tumor.

6. Use of the human breast malignant phyllodes tumor cell line according to claim 1 in screening drug targets for human breast malignant phyllodes tumor.

7. Use of the human breast malignant phyllodes tumor cell line according to claim 1 in screening or evaluating anti-human breast malignant phyllodes tumor drug agents.

8. A cell model, characterized in that The cell model is the cell line of human breast malignant phyllodes tumor according to claim 1.

9. A method for constructing the human breast malignant phyllodes tumor cell line according to claim 1, characterized in that: The steps include: (1) Human breast malignant phyllodes tumor tissue was inoculated into the mammary fat pad of immunodeficient mice to construct a mouse xenograft tumor model derived from patient tissue; (2) After culturing the mice, the tumor tissue was removed; (3) inoculating the tumor tissue block from step (2) into the mammary fat pad of immunodeficient mice to perform passage of the mouse xenograft tumor model derived from the patient's tissue; (4) After five passages of the patient-derived mouse xenograft tumor model, tumor tissue was removed for primary cell culture; (5) When the cells in step (4) grow to 80% conjugation, cell subculture is performed; (6) After the 50th generation of the step (5), the human breast malignant phyllodes tumor cell line is obtained.

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