Application of bifidobacterium adolescentis in preparation of medicine for treating prostatic cancer

Abstract

The invention discloses application of bifidobacterium adolescentis in preparation of a medicine for treating prostatic cancer, and belongs to the field of biological medicine. According to the application, it is found for the first time that an endogenous fibroblast growth factor 21 (FGF21) has the effect of inhibiting the growth of the prostatic cancer tumor, it is found that the abundance of bifidobacterium adolescentis in FGF21 transgenic mouse intestinal flora is remarkably improved, and the bifidobacterium adolescentis is selected as potential probiotics for inhibiting the growth of the prostatic cancer tumor. The intestinal colonization experiment result shows that the bifidobacterium adolescentis can efficiently inhibit the growth of the prostatic cancer tumor by inhibiting the proliferation of tumor cells, and can be prepared into the medicine for treating the prostatic cancer. The invention provides a new treatment strategy and medicine for the treatment of the prostatic cancer, and also provides a brand new medicine development path and a clinical application scheme for the micro-ecological treatment of the prostatic cancer.

Description

TECHNICAL FIELD

[0001] The application relates to the field of biological medicine, in particular to application of Bifidobacterium adolescentis in preparation of a medicine for treating prostate cancer. BACKGROUND

[0002] Prostate cancer (PCa) is the most common malignancy in some countries and the second leading cause of cancer-related death in men. With the aging of the population, the incidence and mortality of prostate cancer in various countries are on the rise. Most patients are in the early stage of androgen-dependent type, and the prognosis is good, but the disease often progresses to androgen-independent type with high malignancy and treatment resistance, and currently there is a lack of effective control means. In addition to traditional risk factors such as genetics and environment, recent studies have gradually revealed that intestinal microbiota plays a key role in the occurrence, development and treatment response of prostate cancer, but how to effectively intervene in prostate cancer by regulating the flora still needs further exploration.

[0003] Bifidobacterium adolescentis is widely distributed in the intestines of animals and humans and plays an important role in maintaining the health of the host. In the past, it has been commonly used for antioxidant, anti-aging and treatment of intestinal-related diseases such as diarrhea, constipation and enteritis. In recent years, studies have also reported the application of Bifidobacterium adolescentis in refreshing the mind, treating non-alcoholic fatty liver and treating systemic lupus erythematosus. However, there is no report on the research of Bifidobacterium adolescentis in inhibiting the occurrence and development of cancer. Therefore, it is urgent to explore the function of Bifidobacterium adolescentis in treating prostate cancer. SUMMARY

[0004] The application aims to provide application of Bifidobacterium adolescentis in preparation of a medicine for treating prostate cancer, so as to solve the problems in the prior art. Bifidobacterium adolescentis can efficiently inhibit the growth of prostate cancer by inhibiting the proliferation of tumor cells, and can be made into a medicine for treating prostate cancer. The application provides a new treatment strategy and medicine for the treatment of prostate cancer, and also provides a new drug development path and clinical application scheme for the microecological treatment of prostate cancer.

[0005] To achieve the above-mentioned purpose, the application provides the following scheme.

[0006] The application provides application of Bifidobacterium adolescentis in preparation of a medicine for treating prostate cancer.

[0007] Further, the treatment of prostate cancer includes inhibition of prostate cancer cell proliferation.

[0008] Further, the medicine is a medicine for increasing the abundance of Bifidobacterium adolescentis in the intestinal flora.

[0009] The application also provides a use of a reagent for detecting the abundance of Bifidobacterium adolescentis in feces in the preparation of a product for predicting or assisting in predicting the prognosis of prostate cancer.

[0010] The application also provides a product for predicting or assisting in predicting the prognosis of prostate cancer, which comprises a reagent for detecting the abundance of Bifidobacterium adolescentis in feces.

[0011] The application discloses the following technical effects:

[0012] The application first discovers that endogenous fibroblast growth factor 21 (FGF21) has the effect of inhibiting the growth of prostate cancer tumors, and discovers that the abundance of Bifidobacterium adolescentis in the intestinal flora of FGF21 transgenic mice is significantly increased, and Bifidobacterium adolescentis is selected as a potential probiotic for inhibiting the growth of prostate cancer tumors. The results of intestinal colonization experiments show that Bifidobacterium adolescentis can efficiently inhibit the growth of prostate cancer tumors by inhibiting tumor cell proliferation, and can be made into a drug for treating prostate cancer. The application provides a new treatment strategy and drug for the treatment of prostate cancer, and also provides a brand-new drug development path and clinical application scheme for the microecological treatment of prostate cancer. BRIEF DESCRIPTION OF DRAWINGS

[0013] In order to more clearly illustrate the technical solutions in the embodiments of the application or the prior art, the following will briefly introduce the drawings needed in the embodiments. Obviously, the drawings in the following description only constitute some embodiments of the application, and for those skilled in the art, other drawings can also be obtained without creative labor.

[0014] Figure 1 FGF21 limits prostate cancer tumor progression and tumor-related inflammation experimental results; wherein, A is the appearance of the tumor; B is the amount of Ki-67 positive cells in the tumor tissue; C is the amount of IL-6 positive cells in the tumor tissue;

[0015] Figure 2 The experimental results of the composition of the intestinal flora of mice and the tumor growth inhibition of Bifidobacterium adolescentis are shown in the figure; wherein, A is the statistical results of the abundance of the intestinal flora of FGF21 transgenic mice; B is the tumor volume; C is the tumor weight; D is the amount of PCNA positive cells in the tumor tissue;

[0016] Figure 3 The results of the correlation between the abundance of Bifidobacterium adolescentis and the content of tumor inflammatory factor IL-6 in the prostate cancer clinical cohort are shown in the figure. DETAILED DESCRIPTION

[0017] The detailed description set forth below is intended as a description of various example embodiments of the application and is not intended to represent the only embodiments in which the application can be practiced. The term "exemplary" used throughout this description means "serving as an example, instance, or illustration," and should not necessarily be construed as preferred or advantageous over other examples. The detailed description includes specific details for the purpose of providing a thorough understanding of the

[0018] It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the application. Additionally, for a range of values of a parameter, unless otherwise indicated, each intervening value by each intervening value, as well as any other stated or intervening value in that stated range is encompassed. The upper and lower limits of these intervening values can independently be included or excluded in the range, and each smaller range that "falls" within the ambit of the larger range is also encompassed. These smaller ranges can be independently combined with one another into other smaller ranges within the larger ranges.

[0019] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. Although preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can be used in the practice of the present application. All documents mentioned herein are incorporated by reference to disclose and describe the methods and / or materials in connection with which the documents are cited. In case of conflict between the present specification and any document incorporated by reference, the present specification will control.

[0020] Various modifications and changes can be made to the specific embodiments of the application described herein without departing from the scope or spirit of the application. Other embodiments of the application will be apparent to those of ordinary skill in the art from the description and examples presented herein. The description and examples are illustrative of the application and are not intended to limit the scope of the application.

[0021] As used herein, the terms "comprises", "comprising", "includes", "including", "has", "having", "contains", "containing", or variations thereof, are intended to be open-ended terms that mean including, but not limited to.

[0022] The FGF21 transgenic mice of the present application have been disclosed in the literature "Huang X, Yu C, Jin C, et al. Forced expression of hepatocyte-specific fibroblast growth factor 21 delays initiation of chemically induced hepatocarcinogenesis. Mol Carcinog. 2006; 45(12):934-942." The applicant hereby commits to make this biological material available to the public, 20 years from the date of filing of this application.

[0023] The Bifidobacterium adolescentis of this invention has the depositary number ATCC 15703 and was purchased from Mingzhou Biotechnology, product number B81568.

[0024] The Bifidobacterium animalis of this invention has the depositary number ATCC25527 and was purchased from Mingzhou Biotechnology, product number B86059.

[0025] Example 1

[0026] 1. FGF21 Functional Experiment

[0027] 1.1 Experimental Methods

[0028] Using RM-1 prostate cancer cells, tumor-bearing mice (5 × 10⁶) were injected orally into the prostate gland of wild-type mice and FGF21 transgenic mice. 5 (1 cell / animal), tumor tissue was removed 14 days later and the tumor volume was recorded.

[0029] Immunohistochemical experiments were performed on tumor tissue to analyze the number of Ki-67 positive cells and IL-6 positive cells, which are related to cell proliferation.

[0030] 1.2 Experimental Results

[0031] like Figure 1 As shown in Figure A, the tumor volume of FGF21 transgenic mice (FGF21 TG) was significantly smaller than that of wild-type mice (WT), indicating that increasing the expression level of FGF21 can effectively inhibit tumor growth.

[0032] The experimental results of Ki-67 positive cell quantity are as follows: Figure 1 As shown in Figure B, the number of Ki-67 positive cells in prostate cancer tumors of FGF21 transgenic mice (FGF21 Tg) is significantly lower than that of wild-type mice (WT); the experimental results for the number of IL-6 positive cells are as follows. Figure 1 As shown in Figure C, the number of IL-6 positive cells in prostate cancer tumors of FGF21 transgenic mice (FGF21 Tg) is significantly less than that of wild-type mice (WT); indicating that FGF21 has the function of inhibiting cell proliferation in tumors.

[0033] 2. Analysis of mouse gut microbiota

[0034] 2.1 Experimental Methods

[0035] A spontaneous prostate cancer model mouse overexpressing FGF21 was constructed using genetic manipulation. Fresh fecal samples were collected from this genotype and TRAMP mice, and 16S rRNA deep sequencing was performed. LDA Effect Size (LEfSe) multilevel discriminant analysis was used to screen out healthy group-specific enriched strains.

[0036] 2.2 Experimental Results

[0037] The results are as follows Figure 2 As shown in Figure A, the abundance of Bifidobacterium (g_Bifidobacterium) and Bifidobacteriaceae (f_Bifidobacteriaceae) in the fecal samples of FGF21-overexpressing model mice (TRAMP / F21tg) was significantly higher than that in model mice (TRAMP); suggesting that the inhibitory effect of FGF21 overexpression on prostate cancer proliferation is related to the enrichment of Bifidobacterium.

[0038] 3. Functional experiment of Bifidobacterium adolescentis

[0039] 3.1 Experimental Methods

[0040] C57 / BL6 mice were divided into four groups. One group was treated with PBS by gavage as a control group. The other three groups were treated with combined antibiotics to clear the intestinal flora (mice were administered 33.2 mg ampicillin, 33.2 mg neomycin, 33.2 mg metronidazole, and 16.7 mg vancomycin daily by gavage for 5 days). Two of the three groups were then colonized with single strains (Bifidobacterium adolescentis and Bifidobacterium animalis by gavage, 1×10⁻⁶). 10 (CFU / animal / day, administered by gavage for 2 days) to establish a gut microbiota-specific reintegration model, with one group serving as a gut microbiota blank control.

[0041] Using RM-1 prostate cancer cells, four groups of mice underwent orthotopic injection of tumor-bearing cells into the prostate (5 × 10⁻⁶ cells per ... 5 (1 cell / animal) The orthotopic tumor was removed 14 days after inoculation, and the volume and weight of the tumor were measured. Immunohistochemical experiments were performed on the tumor tissue to analyze the number of PCNA-positive cells related to cell proliferation.

[0042] 3.2 Experimental Results

[0043] like Figure 2As shown in Figures B and C, the prostate cancer tumor volume and weight of mice with cleared gut microbiota (Abx) were significantly larger than those of the control group (PBS), indicating that gut microbiota can inhibit prostate cancer growth. The prostate cancer tumor volume and weight of mice specifically replenished with Bifidobacterium adolescentis (B. adolescentis) were significantly or extremely significantly smaller than those of mice with cleared gut microbiota (Abx), and similar to those of the control group (PBS), indicating that Bifidobacterium adolescentis effectively inhibits prostate cancer growth. The prostate cancer tumor volume and weight of mice specifically replenished with Bifidobacterium animalis (B. animalis) were significantly or extremely significantly larger than those of the control group (PBS) and mice specifically replenished with Bifidobacterium adolescentis (B. adolescentis), and similar to those of mice with cleared gut microbiota (Abx), indicating that Bifidobacterium animalis does not have the function of inhibiting prostate cancer growth.

[0044] like Figure 2 As shown in Figure D, the number of PCNA-positive cells in mice specifically replenished with Bifidobacterium adolescentis (B. adolescentis) was the lowest, significantly less than that in the control group mice (PBS); while the number of PCNA-positive cells in mice with gut microbiota clearance (Abx) and mice specifically replenished with Bifidobacterium animalis (B. animalis) was significantly higher than that in the control group mice (PBS), indicating that Bifidobacterium adolescentis can inhibit the proliferation of prostate cancer tumor cells.

[0045] The above results indicate that Bifidobacterium adolescentis has a unique function of inhibiting the proliferation of prostate cancer tumor cells and inhibiting the growth of prostate cancer.

[0046] 4. Correlation analysis between Bifidobacterium adolescentis and IL-6 protein levels

[0047] This invention collected fecal samples from 20 prostate cancer patients and healthy individuals for 16S rRNA deep sequencing, analyzed the relative abundance of Bifidobacterium adolescentis in the fecal samples, and performed correlation analysis with the serum IL-6 protein level at patient admission. The results are as follows: Figure 3 As shown, the data indicate that the abundance of Bifidobacterium adolescentis is significantly negatively correlated with the level of IL-6 protein (P<0.05).

[0048] The embodiments described above are merely preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Various modifications and improvements made by those skilled in the art to the technical solutions of the present invention without departing from the spirit of the present invention should fall within the protection scope defined by the claims of the present invention.

Claims

1. The application of a type of Bifidobacterium adolescentis in the preparation of a drug for treating prostate cancer.

2. The application as described in claim 1, characterized in that, The treatment for prostate cancer includes inhibiting the proliferation of prostate cancer cells.

3. The application as described in claim 1, characterized in that, The drug is used to increase the abundance of Bifidobacterium adolescentis in the intestinal flora.

4. The application of a reagent for detecting the abundance of Bifidobacterium adolescentis in feces in the preparation of products for predicting or assisting in the prediction of prostate cancer prognosis.

5. A product for predicting or assisting in the prediction of prostate cancer prognosis, characterized in that, The product includes reagents for detecting the abundance of Bifidobacterium adolescentis in feces.

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