Application of INPP5F protein in preparation of breast cancer diagnosis and / or prognosis product

Abstract

The invention belongs to the technical field of biological medicine, and particularly relates to application of INPP5F protein in preparation of breast cancer diagnosis and / or prognosis products. The amino acid sequence of the INPP5F protein is as shown in SEQ ID NO. 1. Experiments show that the expression of the INPP5F protein in breast cancer tissues is obviously higher than that in adjacent para-carcinoma tissues, the INPP5F protein can be used as a biomarker for breast cancer diagnosis, and the expression of the INPP5F protein is closely related to TNM staging and lifetime of breast cancer patients. The method is simple and easy to implement, the diagnosis process is safe and effective, the method is easily accepted by patients, the diagnosis standard is uniform, the influence of personal subjective factors is small, and it is indicated that the NPP5F protein has important significance on prognosis judgment of breast cancer.

Description

Technical Field

[0001] The present application belongs to the field of biomedicine technology, and specifically relates to the use of INPP5F protein in the preparation of breast cancer diagnosis and / or prognosis products. Background Art

[0002] Breast cancer is one of the malignant tumors that seriously affects women's health. Global cancer statistics show that the incidence and mortality rates of breast cancer in women are among the highest worldwide, making it one of the leading causes of cancer death in women. Notably, cancer statistics released by the International Agency for Research on Cancer (IARC) and the National Cancer Center of my country both show that breast cancer is the most common cancer in women in my country, with the highest incidence and mortality rates among female malignant tumors, posing a serious threat to women's health. Currently, the diagnosis of breast cancer relies primarily on clinical breast examinations, mammography, ultrasound, and magnetic resonance imaging. Exploring new diagnostic and prognostic markers for breast cancer will facilitate early detection and diagnosis of breast cancer, promote early treatment, increase the chances of successful treatment, and thus reduce mortality. Summary of the Invention

[0003] The purpose of the present invention is to address the deficiencies of the prior art and provide the use of INPP5F protein in the preparation of breast cancer diagnosis and / or prognosis products, specifically adopting the following technical solutions: In a first aspect, the present invention provides a use of an INPP5F protein in preparing a breast cancer diagnosis and / or prognosis product, wherein the amino acid sequence of the INPP5F protein is shown in SEQ ID NO.1.

[0004] SEQ ID NO.1:

[0005] The INPP5F protein, short for inositol polyphosphate-5-phosphatase F, is located on the long arm of human chromosome 10. The INPP5F gene encodes an 1,132-amino acid inositol 1,4,5-triphosphate (InsP3)-5-phosphatase. The protein contains a 352-amino acid SAC phosphatase domain, which is the primary region of enzymatic activity. The primary function of the INPP5F protein is to degrade phosphatidylinositol 4,5-bisphosphate (PIP2) and phosphatidylinositol 3,4,5-trisphosphate (PIP3) by removing the 5' phosphate group on the inositol ring, thereby regulating cellular function.

[0006] As a further preferred embodiment, the product includes a chip, a kit or a test paper.

[0007] As a further preferred embodiment, the chip includes a gene chip and a protein chip.

[0008] As a further preferred embodiment, the gene chip includes nucleotide probes for detecting the transcription level of the INPP5F gene; and the protein chip includes a specific binding reagent for the INPP5F protein.

[0009] In the present invention, the samples for diagnosis and / or prognosis are derived from breast cancer tissue and / or adjacent cancer tissue of breast cancer patients, suspected breast cancer patients, breast cancer susceptible populations, breast cancer high-risk populations or healthy populations.

[0010] In a second aspect, the present invention provides a kit for the diagnosis and / or prognosis of breast cancer, comprising a detection reagent for detecting a marker of the above-mentioned INPP5F protein; the marker is any one of a radioactive isotope, a nucleotide chromophore, an enzyme, a fluorescent molecule, a chemiluminescent moiety, a magnetic particle, and a bioluminescent moiety.

[0011] As a further preferred embodiment, the radioactive isotope comprises 32 P. 35 S or 125 I; The nucleotide chromophore includes pyrene, perylene or naphthamide; the enzyme includes horseradish peroxidase or alkaline phosphatase; the fluorescent molecule includes fluorescein isothiocyanate, rhodamine, Cy3 or Cy5; the chemiluminescent part includes acridinium ester, luminol or a luminol derivative; the bioluminescent part includes biotin or digoxigenin.

[0012] As a further preferred embodiment, the detection reagent is used to detect markers of the INPP5F protein by methods including radiometric methods, immunological methods, fluorescence methods, flow cytometry, latex turbidimetry, biochemical methods, enzymatic methods, hybridization methods, gas chromatography-mass spectrometry, liquid chromatography-mass spectrometry, chromatography, chemiluminescence methods, magnetoelectric methods, or photoelectric conversion methods.

[0013] In a third aspect, the present invention provides an application of an INPP5F protein as a target in screening and preparing breast cancer drugs, wherein the amino acid sequence of the INPP5F protein is shown in SEQ ID NO.1.

[0014] In a fourth aspect, the present invention provides an application of an INPP5F protein in constructing a breast cancer prognosis prediction model, wherein the amino acid sequence of the INPP5F protein is shown in SEQ ID NO.1.

[0015] In a fifth aspect, the present invention provides a use of an INPP5F protein in the preparation of a breast cancer tumor suppressor, wherein the amino acid sequence of the INPP5F protein is shown in SEQ ID NO.1.

[0016] The beneficial effects of the present invention are: (1) The present invention provides an INPP5F protein that can be used for the diagnosis or prognosis of breast cancer. Experiments according to the present invention show that the expression of INPP5F protein in breast cancer tissue is significantly higher than that in adjacent paracancerous tissue, and can be used as a biomarker for the diagnosis of breast cancer. In addition, it is shown that the expression of INPP5F protein is closely related to the TNM stage and survival of breast cancer patients.

[0017] (2) During the experimental process of the present invention, immunohistochemical staining was used to detect samples, and it was determined that INPP5F protein can be used as a molecular marker for diagnosing breast cancer. The method is simple and easy to use, the diagnostic process is safe and effective, and it is easily accepted by patients. The diagnostic criteria are unified and less affected by personal subjective factors. INPP5F protein is of great significance for the prognosis of breast cancer. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the following briefly introduces the drawings required for use in the embodiments. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0019] Figure 1 The results show the comparison of INPP5F expression in breast cancer and adjacent adjacent tissues; ***Statistically significant (p < 0.001); Figure 2 Shown is the correlation between INPP5F expression and prognosis of breast cancer patients, p < 0.001; Figure 3 Shown are the ROC analysis results of INPP5F in predicting breast cancer occurrence. DETAILED DESCRIPTION

[0020] The following will be combined with the drawings in the embodiments of this application to clearly and completely describe the technical solutions in the embodiments of this application. Obviously, the embodiments described are part of the embodiments of this application, not all of them. Based on the embodiments in this application, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of this application.

[0021] Breast cancer is one of the most serious malignant tumors affecting women's health. Currently, breast cancer diagnosis relies primarily on clinical breast examinations, mammography, ultrasound, and magnetic resonance imaging. Exploring new diagnostic and prognostic markers for breast cancer will facilitate early detection and diagnosis, promote early treatment, increase the chances of successful treatment, and thus reduce mortality.

[0022] The INPP5F protein, short for inositol polyphosphate-5-phosphatase F, is located on the long arm of human chromosome 10 and plays a potential regulatory role in various disease models. However, the expression and role of INPP5F in breast cancer remain unclear.

[0023] Based on this, the present invention provides the use of an INPP5F protein in the preparation of a breast cancer diagnosis and / or prognosis product. The amino acid sequence of the INPP5F protein is shown in SEQ ID NO. 1. In the present invention, immunohistochemical staining is used to detect the protein expression score of the INPP5F protein in a test sample. Patients with an INPP5F protein expression score greater than 4.5 in the test sample are diagnosed as having breast cancer or at high risk of breast cancer, which is of great significance for the diagnosis and prognosis of breast cancer.

[0024] Example 1 Analysis of NPP5F protein expression in breast cancer and adjacent tissues 1. Source of tissue samples: 172 breast cancer patient samples were collected from the tissue sample library of Shanghai Zhuohao Pharmaceutical Technology Co., Ltd. Surgery dates ranged from January 2004 to August 2017, with final follow-up in April 2022. All patients were pathologically confirmed to have breast cancer. All patients were female, ranging in age from 31 to 84 years, with a median age of 48 years. Of these 172 samples, 130 included both cancer tissue and matched adjacent paracancerous tissue within 1.5 cm of the cancer, while the remaining 42 contained only cancer tissue. Data for these 172 patients are shown in Table 1.

[0025] Table 1 Clinicopathological data of 172 breast cancer patients 2. Tissue microarray production: Tissue microarrays were fabricated by Shanghai Zhuohao Pharmaceutical Technology Co., Ltd. All donor tissue blocks were routinely sectioned and stained with hematoxylin and eosin (HE) staining. A pathologist performed a secondary diagnosis and marked typical pathological sites on the HE sections. Using a Beecher Instruments Inc. tissue microarray machine, 1.5 mm diameter holes were punched into blank recipient wax blocks. Target tissue cores were then obtained from the corresponding locations on the donor tissue blocks according to the markings on the HE slides and placed into the arrayed wells of the recipient blocks. This process was repeated to create the BRC1601 and BRC1603 dot arrays, which matched breast cancer tissue to adjacent adjacent tissues. Serial sections were cut at 4 μm thickness using a paraffin microtome (Leica, Germany) and mounted on imported slides treated to prevent shedding.

[0026] 3. Immunohistochemical staining experiment, the method is as follows: Immunohistochemistry experiments were performed on the prepared tissue chips using the two-step immunohistochemistry kit produced by Beijing Zhongshan Jinqiao Biotechnology Co., Ltd.

[0027] (1) Dewaxing of breast cancer tissue chips: Place the breast cancer tissue chips on a baking machine and bake them at 65°C for two hours. Dewax the tissue chips in a fume hood.

[0028] (2) After dewaxing, the breast cancer tissue chip was rinsed in tap water for 5 minutes; then placed in an immunohistochemistry box containing ddH2O and washed on a shaker for 5 minutes.

[0029] (3) Antigen repair: Add antigen repair solution to the immunohistochemistry box, and then place the breast cancer tissue chip in it; adjust the temperature of the constant temperature water bath to 99°C, place the immunohistochemistry box with the breast cancer tissue chip in the water bath, and use a thermometer to measure the real-time temperature in the immunohistochemistry box. When the temperature rises to 95°C, start timing, and the antigen repair is 18 minutes.

[0030] (4) After antigen retrieval is completed, remove the immunohistochemistry box from the water bath and allow the breast cancer tissue chip to return to room temperature.

[0031] (5) After the breast cancer tissue chip returns to room temperature, add PBST solution to the immunohistochemistry box, place the breast cancer tissue chip in it, and place it on a shaker to wash three times, each time for 5 minutes.

[0032] (6) Removal of peroxidase: Take the breast cancer tissue chip out of the immunohistochemistry box and wipe off the liquid around the tissue with absorbent paper; use an immunohistochemistry pen to draw a circle around the tissue and enclose the tissue in the circle; then place the breast cancer tissue chip in a wet box, add 3% hydrogen peroxide solution to the tissue, and incubate in the dark for 10 minutes.

[0033] (7) After the peroxidase is removed, add PBST solution to the immunohistochemistry box, place the breast cancer tissue chip in it, and place it on a shaker to wash three times, each time for 10 minutes.

[0034] (8) Blocking: Take the breast cancer tissue chip out of the immunohistochemistry box, shake off the liquid on the slide, place the breast cancer tissue chip in a wet box, add blocking liquid to the tissue, cover the wet box, and block at room temperature for one and a half hours.

[0035] (9) Preparation of primary antibody: Prepare INPP5F primary antibody at a ratio of 1:200 using antibody diluent.

[0036] (10) Incubation of primary antibody: After blocking, shake off the blocking solution on the tissue, place the breast cancer tissue chip in a humidified box, add the freshly prepared primary antibody solution to the tissue, cover it with a lid, and incubate it in a 4°C refrigerator overnight.

[0037] (11) After the primary antibody incubation is completed, take out the wet box from the 4°C refrigerator and place it at room temperature for half an hour to allow the breast cancer tissue chip to return to room temperature.

[0038] (12) After returning to room temperature, add PBST solution to the immunohistochemistry box, place the breast cancer tissue chip in it, and place it on a shaker to wash three times, each time for 10 minutes.

[0039] (13) Incubation of secondary antibody: shake off the PBST liquid on the tissue, place the breast cancer tissue chip in a wet box, add HRP-labeled goat anti-rabbit secondary antibody to the tissue, cover it, and incubate at room temperature for one hour.

[0040] (14) After the secondary antibody incubation is completed, add PBST solution to the immunohistochemistry box, place the breast cancer tissue chip in it, and place it on a shaker to wash three times, each time for 10 minutes.

[0041] (15) Preparation of DAB colorimetric solution: Prepare DAB colorimetric solution at a ratio of 1:20. Keep away from light and use immediately after preparation.

[0042] (16) DAB color development: Shake off the PBST liquid on the tissue, place the breast cancer tissue chip under a microscope, and add DAB color development solution to the tissue for color development. When the specific coloring is obvious and the background is clean, place the breast cancer tissue chip in tap water to stop color development.

[0043] (17) Hematoxylin staining: Place the breast cancer tissue chip in hematoxylin staining solution for 2 minutes and 30 seconds, and then rinse in tap water for 5 minutes.

[0044] (18) Hydrochloric acid alcohol differentiation: Dip the breast cancer tissue chip into hydrochloric acid alcohol, quickly take it out, and rinse it in tap water for 5 minutes.

[0045] (19) Anti-blueing with dilute ammonia water: Place the breast cancer tissue chip in dilute ammonia water for anti-blueing for 1 min 30 s, rinse in tap water for 5 min, and dehydrate the tissue.

[0046] (20) Tissue sealing: After the breast cancer tissue chip is dehydrated, the slide is taken out of the transparent liquid, neutral resin is added to the tissue, and then a coverslip is placed on it.

[0047] 4. Quantitative analysis is as follows: The staining results were photographed, and the positive area and staining intensity within each section were converted into corresponding H-Score values ​​to achieve semi-quantitative analysis of tissue staining. The H-Score ranged from 0 to 8, with a median of 4. Grouping was performed based on the H-Score: those with a score above the median were classified as high-expression, and those with a score below or equal to the median were classified as low-expression.

[0048] 5. Statistical analysis is as follows: The expression of INPP5F protein in breast cancer and adjacent adjacent tissues was analyzed using the t-test. The correlation between INPP5F protein expression and clinical indicators of breast cancer patients was analyzed using the chi-square test. The correlation between INPP5F protein expression and prognosis of breast cancer patients was analyzed using the Kaplan-Meier survival analysis and the log-rank test for univariate analysis of survival. A p-value < 0.05 was considered statistically significant.

[0049] 6. The experimental results are as follows: (1) Analysis of INPP5F protein expression in breast cancer and adjacent tissues.

[0050] Immunohistochemical analysis results Figure 1 As shown in Figure 2, the expression of INPP5F protein in breast cancer tissues was significantly higher than that in adjacent tissues. p <0.001.

[0051] (2) Correlation between INPP5F protein and clinical indicators of breast cancer patients.

[0052] Breast cancer patients were grouped according to factors such as age and TNM stage, and the correlation between these factors and INPP5F protein expression was analyzed using a chi-square test. The results, as shown in Table 2, showed no significant statistical correlation between INPP5F protein expression in breast cancer and patient age (p>0.05), but were strongly correlated with TNM stage (p<0.001).

[0053] Table 2 Correlation between INPP5F protein expression and clinical indicators in breast cancer patients (3) Follow-up of breast cancer patients is as follows: surgery time is from January 2004 to August 2017, and the final follow-up time is April 2022. The univariate analysis of survival time using Kalplan-Meier survival analysis and log-rank statistical test showed that, as Figure 2 As shown in the results, breast cancer patients with low INPP5F expression in their cancer tissues had a longer overall survival (p<0.001), and their 5-year survival rate was significantly higher than that of breast cancer patients with high INPP5F expression. This result suggests that the detection of INPP5F is of great significance for the prognosis of breast cancer.

[0054] Example 2 Validation of NPP5F protein in the diagnosis of breast cancer S1. Tissue sample sources are as follows: 105 breast cancer tissue samples and 105 adjacent tissue samples were obtained from the tissue sample library of Shanghai Xinchao Biotechnology Co., Ltd.

[0055] S2. Tissue microarray production: Tissue microarrays were fabricated by Shanghai Xinchao Biotechnology Co., Ltd. Using the TMAGrand Master tissue microarray fabrication system, target tissue cores were punched into recipient wax blocks. The target tissue cores were then placed into the 1.5 mm diameter array wells in the recipient wax blocks. The above steps were repeated to create the HBrED120CS01 and HBrED090CS01 dot arrays of breast cancer and adjacent paracancerous tissue. Serial sections were sectioned at 4 μm thickness using a paraffin microtome (Leica, Germany) and mounted on imported slides treated to prevent shedding.

[0056] 3. Immunohistochemical staining experiment, the method is as follows: Immunohistochemistry experiments were performed on the prepared tissue chips using the two-step immunohistochemistry kit produced by Beijing Zhongshan Jinqiao Biotechnology Co., Ltd.

[0057] (1) Dewaxing of breast cancer tissue chips: Place the breast cancer tissue chips on a baking machine and bake them at 65°C for two hours. Dewax the tissue chips in a fume hood.

[0058] (2) After dewaxing, the breast cancer tissue chip was rinsed in tap water for 5 minutes; then placed in an immunohistochemistry box containing ddH2O and washed on a shaker for 5 minutes.

[0059] (3) Antigen repair: Add antigen repair solution to the immunohistochemistry box, and then place the breast cancer tissue chip in it; adjust the temperature of the constant temperature water bath to 99°C, place the immunohistochemistry box with the breast cancer tissue chip in the water bath, and use a thermometer to measure the real-time temperature in the immunohistochemistry box. When the temperature rises to 95°C, start timing, and the antigen repair is 18 minutes.

[0060] (4) After antigen retrieval is completed, remove the immunohistochemistry box from the water bath and allow the breast cancer tissue chip to return to room temperature.

[0061] (5) After the breast cancer tissue chip returns to room temperature, add PBST solution to the immunohistochemistry box, place the breast cancer tissue chip in it, and place it on a shaker to wash three times, each time for 5 minutes.

[0062] (6) Removal of peroxidase: Take the breast cancer tissue chip out of the immunohistochemistry box and wipe off the liquid around the tissue with absorbent paper; use an immunohistochemistry pen to draw a circle around the tissue and enclose the tissue in the circle; then place the breast cancer tissue chip in a wet box, add 3% hydrogen peroxide solution to the tissue, and incubate in the dark for 10 minutes.

[0063] (7) After the peroxidase is removed, add PBST solution to the immunohistochemistry box, place the breast cancer tissue chip in it, and place it on a shaker to wash three times, each time for 10 minutes.

[0064] (8) Blocking: Take the breast cancer tissue chip out of the immunohistochemistry box, shake off the liquid on the slide, place the breast cancer tissue chip in a wet box, add blocking liquid to the tissue, cover the wet box, and block at room temperature for one and a half hours.

[0065] (9) Preparation of primary antibody: Prepare INPP5F primary antibody at a ratio of 1:200 using antibody diluent.

[0066] (10) Incubation of primary antibody: After blocking, shake off the blocking solution on the tissue, place the breast cancer tissue chip in a humidified box, add the freshly prepared primary antibody solution to the tissue, cover it with a lid, and incubate it in a 4°C refrigerator overnight.

[0067] (11) After the primary antibody incubation is completed, take out the wet box from the 4°C refrigerator and place it at room temperature for half an hour to allow the breast cancer tissue chip to return to room temperature.

[0068] (12) After returning to room temperature, add PBST solution to the immunohistochemistry box, place the breast cancer tissue chip in it, and place it on a shaker to wash three times, each time for 10 minutes.

[0069] (13) Incubation of secondary antibody: shake off the PBST liquid on the tissue, place the breast cancer tissue chip in a wet box, add HRP-labeled goat anti-rabbit secondary antibody to the tissue, cover it, and incubate at room temperature for one hour.

[0070] (14) After the secondary antibody incubation is completed, add PBST solution to the immunohistochemistry box, place the breast cancer tissue chip in it, and place it on a shaker to wash three times, each time for 10 minutes.

[0071] (15) Preparation of DAB colorimetric solution: Prepare DAB colorimetric solution at a ratio of 1:20. Keep away from light and use immediately after preparation.

[0072] (16) DAB color development: Shake off the PBST liquid on the tissue, place the breast cancer tissue chip under a microscope, and add DAB color development solution to the tissue for color development. When the specific coloring is obvious and the background is clean, place the breast cancer tissue chip in tap water to stop color development.

[0073] (17) Hematoxylin staining: Place the breast cancer tissue chip in hematoxylin staining solution for 2 minutes and 30 seconds, and then rinse in tap water for 5 minutes.

[0074] (18) Hydrochloric acid alcohol differentiation: Dip the breast cancer tissue chip into hydrochloric acid alcohol, quickly take it out, and rinse it in tap water for 5 minutes.

[0075] (19) Anti-blueing with dilute ammonia water: Place the breast cancer tissue chip in dilute ammonia water for anti-blueing for 1 min 30 s, rinse in tap water for 5 min, and dehydrate the tissue. (20) Tissue sealing: After the breast cancer tissue chip is dehydrated, the slide is taken out of the transparent liquid, neutral resin is added to the tissue, and then a coverslip is placed on it.

[0076] 4. Quantitative analysis, the method is as follows: The staining results were photographed, and the positive area and staining intensity within each section were converted into corresponding H-Score values ​​to achieve semi-quantitative analysis of tissue staining. The H-Score ranged from 0 to 8, with a median of 4. Grouping was performed based on the H-Score: those with a score above the median were classified as high-expression, and those with a score below or equal to the median were classified as low-expression.

[0077] 5. Statistical analysis is as follows: The diagnostic efficacy of the protein expression intensity value of INPP5F protein for breast cancer was analyzed using ROC curve analysis.

[0078] 6. The experimental results are as follows: AUC is the area under the ROC curve, which is the most commonly used parameter to evaluate the characteristics of the ROC curve and is also an important indicator of test accuracy. Figure 3 The area under the curve (AUC) was 0.930, with a 95% confidence interval (CI) of 0.894–0.967 and p<0.001, indicating that INPP5F protein can be used as a biomarker for diagnosing breast cancer. When the immunohistochemical score (H-Score) for INPP5F protein was 4.5, the sensitivity was 92.4% and the specificity was 87.6%. When tested individually, patients with an H-Score greater than 4.5 were diagnosed with breast cancer with an accuracy of 89.5%.

[0079] The embodiments of the present application are described above in conjunction with the accompanying drawings. Specific examples are used herein to illustrate the principles and implementation methods of the present application. The description of the above embodiments is only used to help understand the core idea of ​​the present application, but the present application is not limited to the above-mentioned specific implementation methods. The above-mentioned specific implementation methods are merely illustrative and not restrictive. Under the guidance of the present application, ordinary technicians in this field can also make many forms without departing from the scope of protection of the purpose of the present application and the claims, all of which are within the protection of the present application.

Claims

1. A use of INPP5F protein in the preparation of a breast cancer diagnosis and / or prognosis product, characterized in that: The amino acid sequence of the INPP5F protein is shown in SEQ ID NO.

1.

2. The use according to claim 1, characterized in that The product includes a chip, a test kit or a test paper.

3. The use according to claim 2, characterized in that The chips include gene chips and protein chips.

4. The use according to claim 3, characterized in that The gene chip includes a nucleotide probe for detecting the transcription level of the INPP5F gene; and the protein chip includes a specific binding reagent for the INPP5F protein.

5. A kit for diagnosis and / or prognosis of breast cancer, characterized in that: A detection reagent comprising a marker for detecting the INPP5F protein of claim 1; the marker is any one of a radioisotope, a nucleotide chromophore, an enzyme, a fluorescent molecule, a chemiluminescent portion, a magnetic particle, and a bioluminescent portion.

6. The kit according to claim 5, characterized in that The radioactive isotopes include 32 P. 35 S or 125 I; The nucleotide chromophore includes pyrene, perylene or naphthamide; the enzyme includes horseradish peroxidase or alkaline phosphatase; the fluorescent molecule includes fluorescein isothiocyanate, rhodamine, Cy3 or Cy5; the chemiluminescent part includes acridinium ester, luminol or a luminol derivative; the bioluminescent part includes biotin or digoxigenin.

7. The kit according to claim 5, characterized in that The detection reagent is used to detect markers of the INPP5F protein, including radiometric methods, immunological methods, fluorescence methods, flow cytometry, latex turbidimetry, biochemical methods, enzymatic methods, hybridization methods, gas chromatography-mass spectrometry, liquid chromatography-mass spectrometry, chromatography, chemiluminescence methods, magnetoelectric methods, or photoelectric conversion methods.

8. A method for using INPP5F protein as a target in screening and preparing breast cancer drugs, characterized in that: The amino acid sequence of the INPP5F protein is shown in SEQ ID NO.

1.

9. A use of INPP5F protein in constructing a breast cancer prognosis prediction model, characterized in that: The amino acid sequence of the INPP5F protein is shown in SEQ ID NO.

1.

10. A use of INPP5F protein in the preparation of a breast cancer tumor suppressor, characterized in that: The amino acid sequence of the INPP5F protein is shown in SEQ ID NO.1.

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