Application of ganoderma lucidum exosome or composition thereof in preparation of anti-breast cancer product

Abstract

The invention particularly relates to application of a ganoderma lucidum exosome or a composition thereof in preparation of an anti-breast cancer product. The invention proves that the ganoderma lucidum exosome can be taken by breast cancer cells and exerts in-vitro activity of inhibiting proliferation and migration of the breast cancer cells and hindering repair of the breast cancer tumor cells, and in an in-vivo disease model, the ganoderma lucidum exosome also exerts the toxicity of the breast cancer tumor cells and effectively inhibits the process of the breast cancer. The research conclusion of the invention provides a development direction of taking the ganoderma lucidum exosome as an anti-breast cancer active component, and the ganoderma lucidum exosome has a good development prospect.

Description

Technical Field

[0001] The present invention relates to the technical field of anti-breast cancer active ingredients, and in particular to the application of Ganoderma lucidum exosomes or a composition thereof in the preparation of anti-breast cancer products. Background Art

[0002] The information disclosed in this background technology section is only intended to enhance the understanding of the overall background of the invention, and should not necessarily be regarded as an admission or any form of suggestion that the information constitutes the prior art already known to a person skilled in the art.

[0003] Lingzhi is a famous medicinal edible fungus in my country. Its active ingredients have multiple functions such as immune regulation, cardiovascular protection, liver protection, blood sugar lowering, and sleep aid. It is widely used in daily health care. Lingzhi has rich active ingredients and has been found to have strong anti-tumor activity in in vivo and in vitro experiments. Its exosomes are a natural extracellular vesicle with a diameter of usually between 30-150nm. They can carry a variety of proteins, RNA, DNA and other substances and participate in intercellular communication. In recent years, it has become a hot spot in malignant tumor research due to its unique bioactive ingredients and tumor regulation potential.

[0004] Many studies have shown that Ganoderma lucidum exosomes can target and regulate apoptosis-related signaling pathways and inhibit tumor cell proliferation by delivering non-coding RNA and functional proteins. At the same time, Ganoderma lucidum and its polysaccharides (peptides) can act on mononuclear macrophages, dendritic cells and other cells in the body, increase the sensitivity of tumor cells to radiotherapy and chemotherapy, increase the level of IFN-γ in the tumor microenvironment, induce tumor cell apoptosis, and ultimately kill tumor cells. The anti-tumor effect of Ganoderma lucidum extracts is relatively well disclosed in the prior art, however, the relevant research on the anti-tumor effect of Ganoderma lucidum exosomes is still relatively blank.

[0005] The inventors believe that exosomes have low immunogenicity and high stability. As a natural delivery tool, they can effectively overcome the limitation that traditional chemical drugs are difficult to penetrate the tumor barrier. Therefore, exploring the anti-tumor activity of Ganoderma lucidum exosomes and clarifying the anti-tumor mechanism of Ganoderma lucidum exosomes are of great significance for promoting the development of anti-tumor drugs and opening up a new direction for the integration of natural drugs and biotechnology. Summary of the invention

[0006] The present invention's research has shown that plant exosomes derived from Ganoderma lucidum have the effects of inhibiting the proliferation and migration of breast cancer cells, reducing the repair ability of tumor cells, etc., and can also effectively inhibit the growth of tumor tissue in tumor models in vivo, confirming the application prospect of Ganoderma lucidum exosomes as anti-breast cancer active ingredients in the preparation of anti-breast cancer related preparations.

[0007] Based on the above technical effects, the present invention provides the following technical solutions: Use of Ganoderma lucidum exosomes, or a composition comprising the Ganoderma lucidum exosomes, in the preparation of an anti-breast cancer product.

[0008] The above-mentioned "Ganoderma lucidum exosomes" are extracellular vesicles or exosome-like nanoparticles secreted by Ganoderma lucidum plant cells, with a particle size of 30-150 nm. The varieties of Ganoderma lucidum are selected from Ganoderma lucidum (red), Ganoderma sinense (purple), Ganoderma tsugae (blue), Ganoderma multiplicatum (yellow), Ganoderma leucocontextum (white), or Ganoderma atrum (black). In a preferred embodiment, the variety of Ganoderma lucidum is Ganoderma lucidum (red).

[0009] In the extraction process of the Ganoderma lucidum exosomes, the filtrate after crushing and impurity removal of fresh Ganoderma lucidum is used as the raw material, and the exosome product is separated from the above filtrate based on the principles of centrifugation, size separation, or precipitation. In a preferred embodiment, Ganoderma lucidum can be further enzymolyzed by enzymatic hydrolysis or other means to fully expose plant cells and increase the secretion amount of exosomes.

[0010] For the extraction of the above Ganoderma lucidum exosomes, the present invention provides an embodiment as follows: (1) Grind and juice Ganoderma lucidum soaked in PBS, filter with gauze to obtain a crude filtrate, and centrifuge the crude filtrate at high speed to retain the supernatant; (2) Add a complex enzyme preparation to the supernatant obtained in step (1) for enzymatic hydrolysis to obtain an enzymatic hydrolysate; freeze-centrifuge the enzymatic hydrolysate and filter it through a 100-mesh sieve to obtain a filtrate, and subject the filtrate to tangential flow filtration and concentration to obtain the product.

[0011] In a more specific embodiment, in the above step (1): The soaking time of PBS is 5-7 h.

[0012] The centrifugation speed of the crude filtrate is 800-1200 rpm, and the centrifugation time is 18-22 min.

[0013] In the above step (2): The dosage ratio of the supernatant to the complex enzyme preparation is 0.1-0.3%.

[0014] The complex enzyme preparation is composed of enzymes in the following proportions: 20%wt. pectinase, 40%wt. cellulase, 10%wt. β-1,3-glucanase, 30%wt. ligninase.

[0015] The filter diameter of the tangential flow filtration is 30-600 nm.

[0016] In the above-mentioned "composition containing Ganoderma lucidum exosomes", the Ganoderma lucidum exosomes are used as the active ingredient, and the composition further includes a pharmaceutically acceptable carrier. According to the general understanding in the art, "pharmaceutically acceptable" means that the carrier is chemically and / or physically compatible with other components in the composition, and even has a synergistic effect, and is physiologically compatible with the subjects of the composition or has the effect of preventing, improving or even treating diseases.

[0017] Furthermore, the above-mentioned pharmaceutically acceptable carrier may include, but is not limited to, pharmaceutically acceptable liquids, gels or solid carriers, aqueous media (such as sodium chloride injection, Ringer's injection, isotonic glucose injection, sterile water injection or Ringer's glucose and lactate injection), non-aqueous media (such as non-volatile oils of plant origin, cottonseed oil, corn oil, sesame oil or peanut oil), antimicrobial agents, isotonic agents (such as sodium chloride or dextrose), buffers (such as phosphate or citrate buffer), antioxidants (such as sodium bisulfate), suspension / dispersing agents (such as sodium carboxymethyl cellulose, hydroxypropyl methyl cellulose or polyvinylpyrrolidone), chelating agents (such as EDTA (ethylenediaminetetraacetic acid) or EGTA (ethylene glycol tetraacetic acid)), emulsifying agents (such as polysorbate 80 (Tween 80)), diluents, adjuvants, excipients, auxiliary reagents or a combination of one or more of the above components. Examples of the auxiliary reagents include fillers, binders, disintegrants, buffers, preservatives, lubricants, flavoring agents, thickening agents, coloring agents or emulsifying agents.

[0018] The subjects of the above composition may include mammals or non-mammals. The mammals include mice, livestock, humans, etc. The dosage of the composition varies according to the type, age, gender, weight of the subject, the specific disease or pathological condition to be treated, the severity of the disease or pathological condition, the route of administration and the judgment of the prescribing personnel, which is technical content that can be routinely determined by those skilled in the art.

[0019] The above-mentioned "anti-breast cancer product" includes, but is not limited to, drugs or kits having the functions of detecting, diagnosing, preventing, improving or treating breast cancer diseases.

[0020] The feasible dosage forms of the drug can be used in the form of oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, sprays, etc., external agents, suppositories and sterile injection solutions, and more preferably injections; the feasible administration methods include oral, intravenous, intramuscular, intra-arterial, intramedullary, intrathecal, intracardiac, transdermal, subcutaneous, intraperitoneal, intranasal, intestinal, topical, sublingual or rectal. Oral or non-oral administration is preferred. Among them, "non-oral" includes subcutaneous, intradermal, intravenous, intramuscular, intra-articular, intrasynovial, intrasternal, intrathecal, intralesional and intracranial injection or infusion techniques.

[0021] In the second aspect of the present invention, there is also provided the use of the above Ganoderma lucidum exosomes, or a composition comprising the Ganoderma lucidum exosomes, in the preparation of foods, health products or beverages.

[0022] Examples of the above foods include biscuits, pastries, powder pastes, instant granules, etc.; examples of the health products include tablets, capsules, oral liquids, powders, granules, etc.; examples of the beverages include compound fruit juices, concentrates, carbonated beverages or tea beverages, etc. The above examples are not specific limitations, and any product forms acceptable in the fields of foods, health products or beverages are within the protection scope of the present invention.

[0023] In the above foods, beverages or health products, the dosage of the Ganoderma lucidum exosomes is selected from 0 to 100% (not 0 or 100), and further, it is 10 to 90%, 20 to 80% or 30 to 70%. It should be noted that the foods, beverages or health products described in the above second aspect should not be regarded as an inevitable functional product. Therefore, the Ganoderma lucidum exosomes therein are not necessarily in an active dosage.

[0024] Compared with the prior art, the beneficial effects of the present invention are as follows: The main improvement of the present invention lies in clarifying the anti-breast cancer activity of Ganoderma lucidum exosomes and providing a development direction for Ganoderma lucidum exosomes as anti-breast cancer active ingredients. In the prior art, the anti-tumor activity and mechanism of Ganoderma lucidum extracts have been relatively well studied. The components of Ganoderma lucidum exosomes are quite different from those of the extracts, and their anti-tumor activity still has great unpredictability. However, exosomes themselves have good immunogenicity and stability, and are expected to be used as a by-product of Ganoderma lucidum processing, expanding the development direction of anti-breast cancer drugs and increasing the economic benefits of the Ganoderma lucidum planting industry at the same time. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] The schematic diagrams in the specification, which form a part of the present invention, are used to provide a further understanding of the present invention. The schematic embodiments of the present invention and their descriptions are used to explain the present invention and do not constitute an improper limitation to the present invention.

[0026] Figure 1 It is a process flow chart for the extraction of the Ganoderma lucidum exosomes described in the present invention; Figure 2 It is the particle size distribution and NTA detection results of the Ganoderma lucidum exosomes in Example 1; Figure 2 In which A is the particle size distribution diagram of the Ganoderma lucidum exosomes, Figure 2 In which B is the transmission electron microscope observation photo of the Ganoderma lucidum exosomes; Figure 3 It is the results of CCK8 detection of the viability of breast cancer cells in Example 4; Figure 3 In which A is the picture of breast cancer cells in the blank group and the test group, Figure 3In which, B is the viability value of breast cancer cells in the blank group and the test group; Figure 4 It is the inhibitory result of breast cancer cell proliferation in Example 4; Figure 5 It is the detection result of the anti-tumor effect of Ganoderma lucidum exosomes in vivo in Example 4; Figure 5 In which, A is the picture of tumor-bearing mice in the control group and the test group, Figure 5 In which, B is the picture of the dissected tumor in the control group and the test group; Figure 6 It is the detection result of the anti-breast cancer cell repair effect of Ganoderma lucidum exosomes in Example 4; Figure 7 It is the detection result of the anti-breast cancer cell migration effect of Ganoderma lucidum exosomes in Example 4; Figure 7 In which, A is the crystal violet staining result of breast cancer cells in the blank control group, Figure 7 In which, B is Figure 7 The partial enlarged view of A in which, Figure 7 In which, C is the crystal violet staining result of breast cancer cells in the Ganoderma lucidum exosome test group, Figure 7 In which, D is Figure 7 The partial enlarged view of C in which. Detailed implementation manners

[0027] It should be noted that the following detailed description is illustrative and is intended to provide further explanation of the present invention. Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the technical field to which the present invention belongs.

[0028] It should be noted that the terms used herein are only for describing specific implementation manners and are not intended to limit the exemplary implementation manners of the present invention. As used herein, unless the context clearly indicates otherwise, the singular forms are also intended to include the plural forms. In addition, it should be understood that when the terms "comprising" and / or "including" are used in this specification, they specify the presence of features, steps, operations, devices, components, and / or combinations thereof.

[0029] As introduced in the background art, Ganoderma lucidum, as a medicinal plant, the anti-tumor activity and related mechanisms of its extracts have been relatively fully disclosed in the prior art. Ganoderma lucidum exosomes have been proven to have high biological activity, however, the related research on its anti-tumor activity and mechanisms is still relatively blank. To solve the above technical problems, the present invention has confirmed that Ganoderma lucidum exosomes have multiple inhibitory pathways on breast cancer cells and are expected to be used as an active ingredient for breast cancer treatment in the development of related preparations.

[0030] In order to enable those skilled in the art to more clearly understand the technical solution of the present invention, the following will specifically describe the technical solution of the present invention in detail with reference to specific examples.

[0031] Example 1 In this example, a Ganoderma lucidum exosome is provided, and the extraction method is as follows: (1) Select fresh Ganoderma lucidum for cleaning and preliminary disinfection by irradiation. Select γ-ray as the ray and irradiate for 12 - 14 h at 3 - 5 kGy. Chop the disinfected Ganoderma lucidum to obtain Ganoderma lucidum pieces smaller than 1.5 cm 3 . Immerse the obtained Ganoderma lucidum pieces in PBS for soaking treatment for 6 h. Transfer the soaked Ganoderma lucidum pieces and PBS solution together to a food processor for grinding and juicing, with a rotation speed of 2000 - 4000 rpm / min and a grinding time of 5 - 15 min. Coarsely filter the grinding liquid through three layers of sterile gauze, transfer the obtained coarse filtrate to a 50 mL sterile centrifuge tube for centrifugation to remove impurities, with a centrifugation speed of 1000 rpm and a centrifugation time of 20 min, and retain the supernatant.

[0032] (2) Add a complex enzyme preparation to the supernatant obtained in step (1) for enzymatic hydrolysis treatment to obtain an enzymatic hydrolysate; the enzymatic hydrolysis temperature is 40 - 50 °C, and the enzymatic hydrolysis time is 4 - 8 h; the dosage ratio of the supernatant to the complex enzyme preparation is 0.1 - 0.3%. The above complex enzyme preparation is composed of enzymes in the following proportions: 20%wt. pectinase, 40%wt. cellulase, 10%wt. β-1,3-glucanase, 30%wt. ligninase.

[0033] (3) Transfer the enzymatic hydrolysate obtained in step (2) to a freezing impurity removal device for freezing centrifugation and filtration to remove impurities, and the filtration screen is 100 mesh. Tangential flow filter the obtained filtrate (the filtration diameter is 30 - 600 nm) and concentrate it to obtain plant exosome-like vesicles, and irradiate with bacteria to obtain the Ganoderma lucidum exosome.

[0034] Perform particle size and electron microscopy analysis on the above Ganoderma lucidum exosome: (1) Particle size analysis Add 1 mL of Ganoderma lucidum exosome suspension to the sample cell, and place the sample cell in NanoSight (Nano-ZS) for test analysis, draw a particle size map, and obtain a particle size distribution diagram as shown in Figure 2 A in it. The average diameter of Ganoderma lucidum exosome is 132.8 nm.

[0035] Table 1 Particle size distribution of Ganoderma lucidum exosome (2) Transmission electron microscope (TEM) analysis For TEM imaging, 30 μL of Ganoderma lucidum exosome suspension was added to the discharged specimen grid, and left horizontally to allow the exosomes to adsorb. Then, the liquid was carefully blotted dry with filter paper. It was stained with 2% phosphotungstic acid dye for 2 min, and the liquid was carefully blotted dry with filter paper. After drying for 30 min, the sample was sent for analysis, observed and photographed by TEM, and the obtained transmission electron microscope micrograph is as shown in Figure 2 shown in B of

[0036] Example 2 In this example, the anti-breast cancer activity of the Ganoderma lucidum exosomes described in Example 1 was verified.

[0037] 1. CCK8 assay for detecting the viability of breast cancer cells MDA-MB-231 cells in the logarithmic growth phase of the breast cancer cell line were taken, and the cell suspension was inoculated at 6000 cells / well in a 96-well cell culture plate and allowed to adhere overnight for 12 h. Blank control group, test group of Ganoderma lucidum exosomes at 2E+9 particles / mL, test group of Ganoderma lucidum exosomes at 4E+9 particles / mL, and test group of Ganoderma lucidum exosomes at 8E+9 particles / mL were set respectively; 9 replicates were set, with 4 wells in each group as parallel controls, and the cells were continuously cultured at 37 °C. Cell viability was detected at 0 h and 48 h respectively; the detection method was CCK8 (10 μL / well), incubated at 37 °C for 4 h, placed in the dark for 30 min, and the OD450 absorbance value was measured with an enzyme-linked immunosorbent assay (ELISA) reader to calculate the cell viability.

[0038] The cell viability results at 48 h are as shown in Figure 3 shown, and the cell viability is the ratio of the test group to the blank control group (%). According to Figure 3 shown in B, the activity of breast cancer cells in the test group of Ganoderma lucidum exosomes at 8E+9 particles / mL decreased significantly, which was 38% of the control group.

[0039] 2. Inhibition of breast cancer cell proliferation MDA-MB-231 cells in the logarithmic growth phase of the breast cancer cell line were taken. Blank control groups with 2000 cells / well, 4000 cells / well, and 6000 cells / well, and test groups with 2000 cells / well + Ganoderma lucidum exosomes, 4000 cells / well + Ganoderma lucidum exosomes, and 6000 cells / well + Ganoderma lucidum exosomes were set respectively, and the concentration of Ganoderma lucidum exosomes was 8E+9 particles / mL; different concentrations of cell suspensions were inoculated in a 6-well cell culture plate and allowed to adhere overnight for 12 h; 2 replicates were set, with 2 wells in each group as parallel controls, and the cells were continuously cultured in an incubator at 37 °C for 14 days. The medium was changed every three days until colony formation; on the 14th day, the cells were stained with 1 mL of 0.1% crystal violet solution for 5 min, rinsed with PBS, and the cell proliferation status was observed under a microscope.

[0040] The crystal violet staining solution can stain the cell nuclei dark purple. According to Figure 4It was shown that obvious purple masses appeared in all three culture dishes of the control group, and as the number of cells increased, the number of masses also increased. In the culture dishes of the Ganoderma lucidum exosome group below, no obvious purple masses were seen. The number of cells was 20.51% of that of the control group, fully demonstrating the inhibitory effect of Ganoderma lucidum exosomes on the proliferation of breast cancer cells.

[0041] 3. Detection of the anti-tumor effect of Ganoderma lucidum exosomes in vivo Four-week-old nude mice were divided into a control group and a Ganoderma lucidum exosome group at 8E+9 particles / mL. There were 5 mice in each group. 200w (100 μL) cells per mouse were inoculated under the armpit to establish an in vivo xenograft model of MDA-MB-231 breast cancer CDX. After confirming tumor formation, intratumoral injection of Ganoderma lucidum exosomes at 1 μg / g / 48 h was initiated within 14 days after tumor formation. The size of the tumor was measured every 7 days and recorded. After 3 - 4 weeks, the mice were photographed, dissected, and the tumors were taken.

[0042] The results were as Figure 5 shown. Solid tumors grew under the armpits of the nude mice in both the control group and the Ganoderma lucidum exosome group. However, the average volume of the solid tumors in the Ganoderma lucidum exosome group was significantly smaller than that in the control group, indicating that Ganoderma lucidum exosomes can also effectively inhibit the growth of breast cancer tumors under in vivo conditions.

[0043] 4. Detection of the anti-breast cancer cell repair effect of Ganoderma lucidum exosomes MDA-MB-231 cells in the logarithmic growth phase of the breast cancer cell line were taken and divided into a blank control group and a Ganoderma lucidum exosome test group. The cell suspension was inoculated in a 6-well plate. The blank control group was added with complete medium, and the test group was inoculated with Ganoderma lucidum exosome solution at a concentration of 8E+9 particles / mL. There were 2 replicates, with 2 wells in each group as parallel controls. The cells were continued to be cultured in an incubator at 37 °C for 24 h. The 6-well plate was taken out and placed in a live cell imager. The blank control group was replaced with basal medium for scratch marking, and the test group still added Ganoderma lucidum exosome solution (8E+9 particles / mL). After 48 h, the data of the live cell imager were statistically analyzed to analyze the repair ability of breast cancer cells.

[0044] The results were as Figure 6 shown. The scratch boundary of the tumor cells in the control group was no longer clear after 48 h, and the tumor cells began to migrate into the scratch area. However, the breast cancer cells in the exosome group showed autolytic changes and a large number of deaths, and there were basically no tumor cells migrating into the scratch area. The above results indicate that Ganoderma lucidum exosomes effectively inhibited the repair effect of breast cancer cells.

[0045] 5. Detection of the anti-breast cancer cell migration effect of Ganoderma lucidum exosomes Take breast cancer cell line MDA-MB-231 in logarithmic growth phase, and set up a blank control group and a Ganoderma lucidum exosome test group respectively. The concentration of Ganoderma lucidum exosomes is 8E+9 particles / mL. In the Transwell chamber, inoculate the cell suspension in the upper chamber, and add complete medium or Ganoderma lucidum exosome solution to the lower chamber until the bottom of the upper chamber is immersed. Set 2 replicates, with 2 wells in each group as parallel controls, and continue to culture in an incubator at 37°C for 24 h. Take out the chamber, rinse it with PBS, stain it with 1 mL of 0.1% crystal violet solution for 5 min, wash it with PBS, and observe the cell migration under a microscope.

[0046] The results are as Figure 7 , Figure 7 shown. The staining of crystal violet in the bottom of the transwell chamber after 24 h of culture is shown. The number of cells passing through the chamber in the exosome group was significantly lower than that in the control group, indicating that Ganoderma lucidum exosomes effectively inhibited the migration of breast cancer cells from the upper chamber to the lower chamber.

[0047] The above are only the preferred embodiments of the present invention and are not intended to limit the present invention. For those skilled in the art, various modifications and changes can be made to the present invention. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included within the protection scope of the present invention.

Claims

1. Use of Ganoderma exosomes or a composition containing the Ganoderma exosomes in the preparation of an anti-breast cancer product.

2. The application according to claim 1, wherein The Ganoderma species is selected from Ganoderma lucidum, Ganoderma sinense, Ganoderma applanatum, Ganoderma multiplicatum, Ganoderma tsugae or Ganoderma atrum.

3. The application according to claim 1, characterized in that, The method for extracting the Ganoderma exosomes is as follows: (1) Grind and juice the Ganoderma soaked in PBS, filter through gauze to obtain a crude filtrate, and centrifuge the crude filtrate at a high speed to retain the supernatant; (2) Add a complex enzyme preparation to the supernatant obtained in step (1) for enzymatic hydrolysis to obtain an enzymatic hydrolysate; freeze-centrifuge the enzymatic hydrolysate and filter through a 100-mesh sieve to obtain a filtrate, and subject the filtrate to tangential flow filtration and concentration to obtain the Ganoderma exosomes.

4. The application according to claim 3, wherein In step (1): the soaking time in PBS is 5-7 h; the centrifugation speed of the crude filtrate is 800-1200 rpm, and the centrifugation time is 18-22 min.

5. The application according to claim 3, characterized in that, In step (2): the dosage ratio of the supernatant to the complex enzyme preparation is 0.1-0.3%; the complex enzyme preparation is composed of enzymes in the following proportions: 20%wt. pectinase, 40%wt. cellulase, 10%wt. β-1,3-glucanase, 30%wt. ligninase; the filtration diameter of the tangential flow filtration is 30-600 nm.

6. The application according to claim 1, wherein In the composition containing Ganoderma exosomes, the Ganoderma exosomes are used as an active ingredient, and the composition further includes a pharmaceutically acceptable carrier; The pharmaceutically acceptable carrier may include, but is not limited to, for example, a pharmaceutically acceptable liquid, gel or solid carrier, aqueous medium, non-aqueous medium, antimicrobial agent, isotonic agent, buffer, antioxidant, suspension / dispersant, chelating agent or EGTA, emulsifier, diluent, adjuvant, excipient, auxiliary reagent or a combination of one or more of the above components; the auxiliary reagent is selected from a filler, binder, disintegrant, buffer, preservative, lubricant, flavoring agent, thickening agent, coloring agent or emulsifier.

7. The application according to claim 1, wherein The subjects of the composition include mammals or non-mammals, and the mammals are selected from mice, livestock or humans; the dosage of the composition varies according to the type, age, sex, weight of the subject, the specific disease or pathological condition to be treated, the severity of the disease or pathological condition, the administration route and the judgment of the prescribing personnel, and belongs to the technical content that can be routinely determined by those skilled in the art.

8. The application according to claim 1, characterized in that, The anti-breast cancer product includes, but is not limited to, a drug or kit having the functions of detecting, diagnosing, preventing, improving or treating breast cancer; the feasible dosage forms of the drug are selected from oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, sprays, external agents, suppositories and sterilized injection solutions, and more preferably an injection.

9. The application according to claim 8, characterized in that, The administration methods of the drug are selected from oral, intravenous, intramuscular, intra-arterial, intramedullary, intrathecal, intracardiac, transdermal, subcutaneous, intraperitoneal, intranasal, enteral, topical, sublingual or rectal; Preferably, it is oral or non-oral administration; the non-oral administration includes subcutaneous, intradermal, intravenous, intramuscular, intra-articular, intrasynovial, intrasternal, intrathecal, intralesional and intracranial injection or infusion techniques.

10. Use of Ganoderma exosomes or a composition containing the Ganoderma exosomes in the fields of food, health products or beverages.

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