Use of serine palmitoyltransferase small subunit a as a biomarker in the preparation of a kit for early diagnosis and / or prognosis prediction of breast cancer

Abstract

This invention provides the application of serine palmitoyltransferase small subunit A (SPTSSA) as a biomarker in the preparation of kits for early diagnosis and / or prognostic prediction of breast cancer, belonging to the field of biomedical technology. Through bioinformatics analysis, histological verification, and serological detection, this invention systematically explores the expression characteristics of SPTSSA in breast cancer and its relationship with the tumor microenvironment, and evaluates its potential application value as a prognostic biomarker for breast cancer, hoping to provide potential targets for personalized treatment of breast cancer. Furthermore, this invention confirms SPTSSA as a novel breast cancer biomarker, which is of great significance for the diagnosis, prognosis, and treatment of breast cancer, providing a theoretical basis for SPTSSA-targeted therapeutic strategies.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, and in particular relates to the application of serine palmitoyltransferase small subunit A as a biomarker in the preparation of kits for early diagnosis and / or prognosis prediction of breast cancer. Background Technology

[0002] Breast cancer (BC) is a condition in which mammary epithelial cells proliferate uncontrollably under the influence of various carcinogenic factors. Breast cancer poses a serious threat to women's health. Although there have been advances in breast cancer diagnosis and treatment, such as early screening, surgery, radiotherapy, chemotherapy, and targeted therapy, high recurrence and metastasis rates continue to contribute to the persistently high mortality rate.

[0003] Traditional diagnostic methods for breast cancer are diverse, with imaging examinations being commonly used. Ultrasound, for example, allows observation of the internal structure of the breast and is relatively simple and non-invasive. However, its resolution for early lesions such as microcalcifications is limited, leading to missed diagnoses. X-ray examinations (such as mammography) can detect early breast lumps and microcalcifications, but their effectiveness in detecting lesions in dense breast tissue may be affected, and they pose certain radiation hazards. Tissue biopsies are also important diagnostic tools, such as fine-needle aspiration biopsy, core needle biopsy, and surgical excision biopsy. While these methods directly obtain lesion tissue for pathological diagnosis with high accuracy, they are invasive procedures, causing patient discomfort. Furthermore, the results depend to some extent on the operator's skill and experience; inaccurate sampling sites can lead to misdiagnosis or an inability to accurately assess the severity of the disease. Moreover, these traditional diagnostic methods are generally not ideal for early detection of micro-lesions, often only revealing abnormalities at a certain stage of tumor development, hindering the early and accurate diagnosis of breast cancer.

[0004] Currently, clinical prognostic assessments of breast cancer primarily rely on pathological staging and certain protein biomarkers. Pathological staging considers factors such as tumor size, lymph node metastasis, and distant metastasis to roughly determine a patient's prognosis. However, this approach is relatively macroscopic and struggles to accurately predict individual recurrence and metastasis risks; patients at the same stage may experience significant differences in subsequent disease progression. At the protein biomarker level, markers like carcinoembryonic antigen (CEA) and carbohydrate antigen 15-3 (CA15-3) can, to some extent, aid in assessing disease progression, monitoring treatment effectiveness, and providing some prognostic indications. However, their accuracy is limited when used alone, resulting in unreliable prognostic predictions. Breast cancer is a highly heterogeneous disease, and existing conventional prognostic assessment methods struggle to comprehensively and accurately predict prognoses for different individuals, failing to adequately meet the clinical need for personalized and precise prognostic assessments. Therefore, finding more effective novel biomarkers to compensate for the shortcomings of existing markers is crucial. Summary of the Invention

[0005] To address the shortcomings of existing research, this invention systematically explores the expression characteristics of SPTSSA in breast cancer and its relationship with the tumor microenvironment through bioinformatics analysis, histological verification, and serological detection, and evaluates its potential application value as a prognostic biomarker for breast cancer, providing potential targets for personalized treatment of breast cancer.

[0006] This invention provides the application of serine palmitoyltransferase small subunit A as a biomarker in the preparation of kits for early diagnosis and / or prognosis prediction of breast cancer.

[0007] Preferably, the kit enables early diagnosis of breast cancer and / or prediction of breast cancer prognosis by detecting the expression level of the small subunit A of serine palmitoyltransferase in the subject's sample.

[0008] Preferably, the sample includes a serum sample.

[0009] Preferably, the serine palmitoyltransferase small subunit A is highly expressed in breast cancer samples, and the expression level of the serine palmitoyltransferase small subunit A is significantly different from that in healthy samples; the expression level of the serine palmitoyltransferase small subunit A is significantly different in samples before and after breast cancer treatment intervention.

[0010] This invention also provides the application of reagents for detecting serine palmitoyltransferase small subunit A in the preparation of kits for early diagnosis and / or prognosis prediction of breast cancer.

[0011] Preferably, the reagent includes a reagent for detecting the expression level of serine palmitoyltransferase small subunit A.

[0012] Preferably, the kit is a kit prepared based on immunoassay technology.

[0013] Preferably, the reagents for detecting the expression level of serine palmitoyltransferase small subunit A include enzyme-linked immunosorbent assay (ELISA) reagents and / or multiplex immunohistochemical assay reagents.

[0014] Preferably, the enzyme-linked immunosorbent assay (ELISA) reagent and / or multiplex immunohistochemical assay reagent include an anti-serine palmitoyltransferase small subunit A antibody.

[0015] Beneficial effects:

[0016] This invention provides the application of serine palmitoyltransferase small subunit A as a biomarker in the preparation of kits for early diagnosis and / or prognostic prediction of breast cancer. Simultaneously, it provides the application of reagents for detecting serine palmitoyltransferase small subunit A in the preparation of kits for early diagnosis and / or prognostic prediction of breast cancer. Through analysis of the Cancer Genome Atlas (TCGA) database, this invention clarifies that SPTSSA mRNA is highly expressed in breast cancer and is closely related to clinical characteristics of breast cancer (such as tumor size, lymph node metastasis, TNM stage) and poor prognosis, and can serve as a potential factor for predicting poor prognosis in breast cancer patients. Furthermore, this invention discovers that SPTSSA participates in regulating the immune landscape of the tumor microenvironment, and its expression is related to various immune cells (such as CD4+). 3+ CD 4+ T cells, CD4 68+ CD 163+ SPTSSA is associated with macrophages and immune checkpoints (PD-1, CTLA-4), suggesting a potential role in tumor immune escape. Furthermore, this invention found elevated SPTSSA protein levels in the serum of breast cancer patients, with significant differences before and after surgery, indicating its potential application value in monitoring tumor burden. In summary, this invention confirms that SPTSSA represents a novel biomarker for breast cancer, with significant implications for its diagnosis, prognosis, and treatment, particularly in the context of immunotherapy, providing a theoretical basis for SPTSSA-targeted therapeutic strategies. Attached Figure Description

[0017] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the accompanying drawings used in the embodiments will be briefly described below.

[0018] Figure 1 This is a schematic diagram of the research methods and technical routes in Example 1;

[0019] Figure 2This is a graph showing the results of analyzing SPTSSA mRNA expression in breast cancer and normal breast tissues using the TCGA database in Example 1; *** indicates P<0.001;

[0020] Figure 3 The correlation results between PD-1 and CTLA-4 expression and SPTSSA levels in the breast cancer tumor area and stroma area in Example 1;

[0021] Figure 4 This is a Kaplan-Meier survival curve analysis result of the overall survival of breast cancer patients in the SPTSSA high and low expression groups in Example 1;

[0022] Figure 5 The figure shows the expression results of SPTSSA protein in serum in Example 1; where (a) shows the expression of SPTSSA protein in the serum of healthy individuals and breast cancer patients before surgery, and (b) shows the expression of SPTSSA protein in the serum of breast cancer patients before and after surgery; * indicates P<0.05;

[0023] Figure 6 The ROC curves for SPTSSA protein in preoperative serum of healthy individuals and breast cancer patients in Example 2 are shown.

[0024] Figure 7 The ROC curves for SPTSSA protein in the preoperative and postoperative serum of breast cancer patients in Example 2 are shown. Detailed Implementation

[0025] This invention provides the application of serine palmitoyltransferase small subunit A as a biomarker in the preparation of kits for early diagnosis and / or prognostic prediction of breast cancer. The serine palmitoyltransferase small subunit A described in this invention has accession number Q969W0 in the UniProt database.

[0026] In one embodiment, the kit achieves early diagnosis and / or prognostic prediction of breast cancer by detecting the expression level of the small subunit A of serine palmitoyltransferase in a subject's sample. In one embodiment, the sample includes, but is not limited to, serum samples. In one embodiment, the reagent used to detect the expression level of the small subunit A of serine palmitoyltransferase is an immunoassay reagent; in another embodiment, the immunoassay reagent includes enzyme-linked immunosorbent assay (ELISA) reagents and / or multiplex immunohistochemistry (mIHC) reagents; in yet another embodiment, the immunoassay reagent is an enzyme-linked immunosorbent assay (ELISA) reagent.

[0027] In one embodiment, the serine palmitoyltransferase small subunit A of the present invention is highly expressed in breast cancer samples and the expression level of the serine palmitoyltransferase small subunit A is significantly different from that in healthy samples; the expression level of the serine palmitoyltransferase small subunit A is significantly different in samples before and after breast cancer treatment intervention; in another embodiment, the significant difference is a significant difference with P < 0.05.

[0028] This invention also provides the application of reagents for detecting serine palmitoyltransferase small subunit A in the preparation of kits for early diagnosis and / or prognostic prediction of breast cancer. As one embodiment, the kit described in this invention is a kit prepared based on immunoassay technology.

[0029] In one embodiment, the reagents of the present invention include reagents for detecting the expression level of serine palmitoyltransferase small subunit A; in another embodiment, the reagents for detecting the expression level of serine palmitoyltransferase small subunit A include enzyme-linked immunosorbent assay (ELISA) reagents and / or multiplex immunohistochemical assay reagents; in yet another embodiment, the ELISA reagents and / or multiplex immunohistochemical assay reagents include anti-serine palmitoyltransferase small subunit A antibody.

[0030] This invention systematically explores the expression characteristics of SPTSSA in breast cancer and its relationship with the tumor microenvironment through bioinformatics analysis, histological verification, and serological detection, and evaluates its potential application value as a prognostic biomarker for breast cancer, providing a potential target for personalized treatment of breast cancer. In other words, this invention confirms that SPTSSA represents a new biomarker for breast cancer, which is of great significance for the diagnosis, prognosis, and treatment of breast cancer, especially in the context of immunotherapy, providing a theoretical basis for treatment strategies targeting SPTSSA.

[0031] To further illustrate the present invention, the application of the serine palmitoyltransferase small subunit A provided by the present invention as a biomarker in the preparation of kits for early diagnosis and / or prognosis prediction of breast cancer is described in detail below with reference to the accompanying drawings and embodiments. However, these descriptions should not be construed as limiting the scope of protection of the present invention.

[0032] Example 1

[0033] The schematic diagram of the research methods and technical routes in this embodiment is shown below. Figure 1 As shown.

[0034] 1. Experimental Materials and Methods

[0035] (1) Data Extraction: Pan-Cancer (pan-oncogene set) was obtained from The Cancer Genome Atlas (TCGA), and the expression data of the ENSG00000165389 (SPTSSA) gene in various samples were extracted. Using the RNA sequencing data, the expression of SPTSSA mRNA in breast cancer tissue and non-tumor tissue was compared, including 1092 breast cancer samples and 113 non-tumor tissue samples.

[0036] (2) Clinical sample collection and processing: From 2010 to 2017, 145 breast cancer specimens were collected from the biobank of the Affiliated Hospital of Nantong University to construct tissue microarrays (TMA). TMA divided the samples into 2 mm cores and obtained clinical pathological annotation information such as molecular subtype, histological classification, and patient demographic characteristics corresponding to the TMA. Samples from patients who received neoadjuvant therapy before surgery were excluded.

[0037] Five mL of fasting venous blood was collected from 60 patients before and after surgical intervention, 30 healthy individuals, and 12 pairs of breast cancer patients before and after surgery. After sedimentation for 6 hours, the supernatant was centrifuged and stored at -80°C. Ethical approval and informed consent from the patients were obtained.

[0038] (3) Antibody sources: Anti-SPTSSA monoclonal antibody: S1N1, an anti-SPTSSA monoclonal antibody disclosed in Chinese patent CN112552409A; anti-cytokeratin (CK) antibody (1:4000, orb69073, Biobyt, UK); anti-CD3 antibody (1:800, 85061s, CST, USA); anti-CD4 antibody (1:400, ab133616, USA); anti-CD8 antibody (1:800, 85336s, CST, USA); anti-CD66b antibody (1:400, arg66287, Arigo Biolaboratories). Corp. (China); Anti-CD20 (1:300; ab78237, Abcam); Anti-CD68 antibody (1:400, 76437s, CST, USA), Anti-CD86 antibody (1:400, orb388891, Biorbyt, UK); Anti-CD163 antibody (1:200, 93498s, CST, USA), Anti-LAMP3 antibody (1:400, orb1433531, Biorbyt, USA), Anti-PD1 antibody (1:200, 13684T, CST, USA), Anti-PD-L1 antibody (1:200, 18616s, CST, USA); Anti-CTLA-4 antibody (1:200, orb527271, Biorbyt, UK).

[0039] Multiple immunohistochemistry (mIHC) and scoring:

[0040] 1) Bake breast cancer TMA slices in an oven at 70℃ for 60 minutes, and bake the slices for 60 minutes.

[0041] 2) Dewaxing with xylene (shaking for 6 min) - 100% alcohol (5 min) - 95% alcohol (5 min) - 70% alcohol (5 min), then hydrating and rinsing with ultrapure water;

[0042] 3) Fix in 10% formalin solution for 10 min, then rinse with ultrapure water;

[0043] 4) Dilute the citric acid tissue antigen repair solution (100×) (Fuzhou Maixin Biotechnology Development Co., Ltd.) with ultrapure water at a ratio of 1:100 and carry out the repair. Cover the container (to prevent liquid evaporation during heating), heat on high (100% power) for 2.5 minutes, then heat on low (20% power) for 15 minutes. After removing the container, open the container and allow it to cool naturally to room temperature. Rinse with ultrapure water first, then rinse with TBST.

[0044] 5) Use an immunoassay pen to circle the sample area on the slide, add blocking solution, block for 10 minutes, and then discard.

[0045] 6) Dilute the anti-SPTSSA monoclonal antibody to 1:800 with primary antibody dilution buffer (multiplex fluorescence immunohistochemistry kit) (Benno Panoramic Biotechnology Beijing Co., Ltd.), immerse the sample area, and incubate overnight at 4°C;

[0046] 7) After recovering the antibody, rinse with TBST for 3×2 min, add secondary antibody enhancement solution (horseradish peroxidase (HRP) conjugated secondary antibody, multiplex fluorescent immunohistochemistry kit) to immerse the sample area, incubate at room temperature for 10 min, and rinse with TBST for 3×2 min.

[0047] 8) Immerse the sample area in 100 μl of 1X dye working solution (diluted 1:100 with signal amplification solution) from the kit, incubate at room temperature in the dark for 10 min, and rinse with TBST for 3 × 2 min.

[0048] 9) If multiple antibodies are incubated (the source of the antibodies is recorded in step (3)), repeat steps 4)-8) to decolorize. If only one antibody is stained, repeat step 4 before mounting. After repair, cool to a constant temperature, rinse with TBST and then rinse with ultrapure water.

[0049] 10) Stain and mount with DAPI working solution (multiplex fluorescence immunohistochemistry kit);

[0050] 11) TMA was imaged using the Vectra 3.0 automated quantitative pathological imaging system, which consisted of a tissue microarray system (UT06, Unitma, South Korea), an inverted microscope (DMIRB, Leica, Germany), and a fully automated microplate reader (SN209941, Bio-Tek, USA). Image analysis and scoring were performed using InForm (Perkin Elmer 2.6.0) software, and a threshold for positive or negative cells was set for each cell. The percentage of cells in each region was calculated and scored (0-100 points).

[0051] (4) Enzyme-linked immunosorbent assay (ELISA): The serum concentration of SPTSSA was detected using an ELISA kit. Reagents, samples and standards were prepared according to the manufacturer's instructions. The double antibody sandwich method was used to improve sensitivity and specificity. SPTSSA antibody was diluted 1:3000 and plated on a 96-well plate. After incubation, washing, addition of sample and biotinylated secondary antibody, TMB substrate was used for color development, and absorbance was measured at 630 nm using a Multiskan Go microplate reader.

[0052] (5) Statistical analysis: Pearson correlation analysis was used to compare the baseline differences in clinicopathological parameters between the low or no SPTSSA expression group and the high expression group; the Kaplan-Meier method was used to assess the differences in overall survival among patients with different SPTSSA mRNA expression levels, and a Cox regression model was established to predict the impact of covariates on survival outcomes; the X-tile tool was used to determine the cutoff value of SPTSSA expression, and the statistical significance was set as P<0.05. All tests were two-tailed tests, and the data were expressed as mean, standard deviation or 95% confidence interval. SPSS 24.0 software was used for statistical calculations.

[0053] 2. Test Results

[0054] (1) Expression levels of SPTSSA mRNA and protein in breast cancer tissues

[0055] TCGA analysis revealed a significant increase in SPTSSA mRNA levels in breast cancer tissues compared to non-tumor tissues (P<0.05), suggesting high expression of SPTSSA in breast cancer tissues. Figure 2 As shown.

[0056] mIHC quantitative analysis based on tissue microarrays constructed from 145 breast cancer specimens showed that SPTSSA protein was mainly positively expressed in breast cancer cell tissues. Compared with peripheral non-tumor tissues, the expression level in tumor tissues was significantly increased, and it was consistent with the mRNA expression pattern in TCGA. Breast cancer patients in the high expression group had a poorer clinical prognosis.

[0057] (2) SPTSSA as an independent factor for survival prediction

[0058] Based on the mIHC measurement results and clinicopathological information of 145 breast cancer specimens (Table 1), SPTSSA was analyzed as an independent factor for survival prediction, and the results are shown in Table 2.

[0059] Table 1. Clinicopathological information of specimens from 1145 breast cancer patients

[0060]

[0061]

[0062] Note: Blank cells in Table 1 indicate no data, and the same applies to Table 2.

[0063] Table 2 shows the results of the independent factor analysis of SPTSSA as a survival predictor.

[0064]

[0065] It can be seen that SPTSSA protein levels are related to tumor size (x). 2 =8.761, P=0.013), degree of lymph node involvement (x 2 =10.713, P=0.005) and TNM staging (x 2 =9.818, P=0.007) significantly correlated with TNM stage, and the protein level of SPTSSA in breast cancer stroma was also significantly correlated with TNM stage (x 2 =7.446, P=0.024).

[0066] Univariate and multivariate Cox regression analyses showed that SPTSSA was significantly associated with TNM stage (P = 0.001), affecting overall survival and serving as an independent prognostic factor.

[0067] (3) The relationship between SPTSSA protein and immune infiltrating cells

[0068] Correlation analysis showed that SPTSSA protein expression was associated with CD68 expression in the tumor region. + CD163 + Macrophages (r = 0.234, P = 0.01) and CD3+ in the stromal region + CD4 + The presence of T cells (r = 0.200, P = 0.018) was significantly associated with this finding.

[0069] Further mIHC analysis showed that the expression of PD-1 (r = 0.220, P = 0.008) and CTLA-4 (r = 0.225, P = 0.007) was positively correlated with SPTSSA levels in breast cancer tumor regions, while no significant correlation was found in the stromal region. The results are as follows: Figure 3 As shown, the left figure shows the correlation between the expression of PD-1 and CTLA-4 and the SPTSSA level in the breast cancer tumor area, and the right figure shows the correlation between the expression of PD-1 and CTLA-4 and the SPTSSA level in the stroma of breast cancer.

[0070] (4) Based on the SPTSSA protein expression measured by mIHC in 145 patients, they were divided into high and low expression groups. Combined with the survival time of these breast cancer patients, Kaplan-Meier survival curves for overall survival were analyzed and plotted. The results are as follows: Figure 4 As shown.

[0071] (5) SPTSSA protein level in patient serum

[0072] Based on the serum ELISA detection and analysis results of 60 patients before and after surgical intervention, it was found that the serum SPTSSA protein level of breast cancer patients was significantly increased compared with that of 30 healthy controls (P<0.05). Based on the serum ELISA detection and analysis results of 12 pairs of breast cancer patients before and after surgery, it was found that the preoperative serum level was higher than the postoperative level (P<0.05). Figure 5 As shown.

[0073] The samples and sources used in step (5) are the same as in Example 2.

[0074] 3. Experimental Conclusions

[0075] (1) SPTSSA is highly expressed in breast cancer tissues and is closely related to the clinical characteristics of breast cancer (such as tumor size, lymph node metastasis, TNM stage, etc.) and poor prognosis. It can be used as a potential factor to predict poor prognosis in breast cancer patients.

[0076] (2) SPTSSA participates in regulating the immune landscape of the tumor microenvironment, and its expression is associated with various immune cells (such as CD3). + CD4 + T cells, CD68 + CD163 + It is associated with macrophages and immune checkpoints (PD-1, CTLA-4), suggesting that it may play a role in tumor immune escape.

[0077] (3) The level of SPTSSA protein in the serum of breast cancer patients is elevated, and there are differences before and after surgery, indicating that it has potential application value in monitoring tumor burden.

[0078] (4) SPTSSA represents a new biomarker that is of great significance for the prognosis and treatment of breast cancer, especially in the context of immunotherapy. The above-mentioned characteristics of SPTSSA provide a theoretical basis for developing treatment strategies that target SPTSSA.

[0079] Example 2

[0080] Sensitivity and specificity of SPTSSA protein as a biomarker for breast cancer diagnosis and prognosis.

[0081] 1. Subject population

[0082] The study population in this embodiment consisted of 30 healthy individuals undergoing routine checkups and 72 breast cancer patients. Preoperative serum samples from the 30 healthy individuals and 60 breast cancer patients were used to assess the sensitivity and specificity of SPTSSA protein as a diagnostic marker for breast cancer. Preoperative and postoperative serum samples from the remaining 12 breast cancer patients were used to assess the sensitivity and specificity of SPTSSA as a prognostic marker. The healthy individuals undergoing routine checkups were from the Health Examination Center of the Affiliated Hospital of Nantong University. All serum samples from breast cancer patients were collected when they were diagnosed with breast cancer but before receiving any radiotherapy, chemotherapy, or surgical treatment, and were stored at -80°C.

[0083] 2. The steps for detecting the sensitivity and specificity of SPTSSA protein in the test population are as follows:

[0084] (1) Coating: Dilute SPTSSA antibody 1:3000 with coating buffer (Beijing Solarbio Technology Co., Ltd.), add 100 μl / well to a 96-well plate, and incubate overnight at 4°C;

[0085] (2) Washing: Discard the liquid in the wells, wash with PBST for 3×3 min, and drain.

[0086] (3) Blocking: Add 200 μl of 1% BSA per well and incubate at 37°C in the dark for 2 hours;

[0087] (4) Discard the liquid in the well, wash with PBST, 3×3min, and drain.

[0088] (5) Sample addition: 100 μl / well, incubate at 37℃ in the dark for 1 h;

[0089] (6) Washing: Discard the liquid in the wells, wash with PBST for 3×3 min, and drain.

[0090] (7) Secondary antibody: diluted with HRP-SPTSSA 1:1000, 100 μl / well, incubated at 37℃ in the dark for 1 h;

[0091] (8) Washing plate: Discard the liquid in the wells, wash with PBST for 3×3 min, and drain.

[0092] (9) Color development: 100 μl of 1×TMB solution (Beijing Solarbio Science & Technology Co., Ltd.) per well, incubate at 37℃ for 1-30 min;

[0093] (10) Termination of reaction: After complete color development, add 50 μl of TMB stop solution (Beijing Solarbio Technology Co., Ltd.) per well to terminate the color development reaction;

[0094] (11) The absorbance was measured at 450 nm using the Thermo Mul-tiskan Go full-wavelength microplate reader.

[0095] (12) Plot the standard curve and analyze the data.

[0096] ROC curve analysis was performed using GraphPadPrism software (version 10.4.0) to evaluate the performance of the prediction model. MedCalc software (version 23.0.2) was used for calculations, and the optimal critical value was determined by maximizing the Youden index (sensitivity + specificity - 1). Sensitivity and specificity were then determined based on the selection of the optimal critical value. The experimental results are shown in Table 3.

[0097] Table 3. Sensitivity and specificity of SPTSSA as a breast cancer marker.

[0098] antigen Sensitivity Specificity SPTSSA (Health and Breast Cancer) 85.00％ 88.33％ SPTSSA (Pre- and Post-operative) 68.52％ 81.48％

[0099] The ROC curve analysis results of SPTSSA protein as a marker for breast cancer diagnosis and prognosis are as follows: Figure 6 and Figure 7 As shown.

[0100] Depend on Figures 6-7 The results showed that the area under the curve (AUC) for SPTSSA protein levels in preoperative serum of healthy individuals and breast cancer patients was 0.9108 (0.8587-0.9629), with a P-value < 0.001; the AUC for SPTSSA protein levels in preoperative serum of breast cancer patients was 0.8014 (0.7204-0.8824), with a P-value < 0.001. Significance was defined as P < 0.05.

[0101] The optimal cutoff, sensitivity, and specificity for SPTSSA protein in predicting breast cancer were as follows: At a cutoff value > 1.6188, the area under the ROC curve (AUC) of SPTSSA protein in preoperative serum of healthy individuals and breast cancer patients was 0.9108 (95% CI: 0.8587–0.9629), with a sensitivity of 85.00% and a specificity of 88.33%, which was statistically significant (P < 0.001). At a cutoff value > 1.7546, the AUC of SPTSSA protein in preoperative serum of breast cancer patients compared to postoperative serum was 0.8014 (95% CI: 0.7204–0.8824), with a sensitivity of 68.52% and a specificity of 81.48%, also statistically significant (P < 0.001).

[0102] It is evident that the ROC curve of SPTSSA protein has good diagnostic value in the comparison of serum levels in healthy individuals and breast cancer patients before and after surgery. Therefore, SPTSSA may serve as a good non-invasive diagnostic and prognostic biomarker for breast cancer.

[0103] Although the above embodiments have provided a detailed description of the present invention, they are only some embodiments of the present invention, and not all embodiments. People can obtain other embodiments based on these embodiments without creative effort, and these embodiments all fall within the protection scope of the present invention.

Claims

1. Application of serine palmitoyltransferase small subunit A as a biomarker in the preparation of kits for early diagnosis and / or prognostic prediction of breast cancer.

2. The application according to claim 1, characterized in that, The kit enables early diagnosis and / or prognostic prediction of breast cancer by detecting the expression level of the small subunit A of serine palmitoyltransferase in subject samples.

3. The application according to claim 2, characterized in that, The samples include serum samples.

4. The application according to claim 2, characterized in that, The small subunit A of serine palmitoyltransferase was highly expressed in breast cancer samples and showed a significant difference in expression level compared to healthy samples; the expression level of the small subunit A of serine palmitoyltransferase was significantly different in samples before and after breast cancer treatment intervention.

5. Application of reagents for detecting serine palmitoyltransferase small subunit A in the preparation of kits for early diagnosis and / or prognosis prediction of breast cancer.

6. The application according to claim 5, characterized in that, The reagents include those for detecting the expression level of the small subunit A of serine palmitoyltransferase.

7. The application according to claim 5 or 6, characterized in that, The kit is prepared based on immunoassay technology.

8. The application according to claim 6, characterized in that, The reagents used to detect the expression level of serine palmitoyltransferase small subunit A include enzyme-linked immunosorbent assay (ELISA) reagents and / or multiplex immunohistochemical assay reagents.

9. The application according to claim 8, characterized in that, The enzyme-linked immunosorbent assay (ELISA) reagents and / or multiplex immunohistochemistry (MHI) reagents include antiserine palmitoyltransferase small subunit A antibody.

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