Application of serine palmitoyl transferase small subunit A as biomarker in preparation of kit for early diagnosis and / or prognosis prediction of breast cancer

Abstract

The invention provides application of a serine palmitoyl transferase small subunit A (SPTSSA) as a biomarker in preparation of a kit for early diagnosis and / or prognosis prediction of breast cancer, and belongs to the technical field of biomedicine. Through bioinformatics analysis, histological verification and serological detection, expression characteristics of the SPTSSA in the breast cancer and the relationship between the SPTSSA and a tumor microenvironment are systematically discussed, the potential application value of the SPTSSA as a breast cancer prognosis marker is evaluated, and a potential target spot is expected to be provided for individualized treatment of the breast cancer. The invention confirms that the SPTSSA is a novel breast cancer biomarker, has important significance for diagnosis, prognosis and treatment of breast cancer, and provides a theoretical basis for a treatment strategy of targeted SPTSSA.

Description

Technical Field

[0001] The invention belongs to the field of biomedical technology, and in particular relates to the 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. Background Art

[0002] Breast carcinoma (BC) is a phenomenon in which mammary epithelial cells proliferate uncontrollably under the influence of multiple carcinogenic factors. Breast cancer seriously threatens women's health. Although there have been advances in the diagnosis and treatment of breast cancer, such as early screening, surgery, radiotherapy, chemotherapy and targeted therapy, the high recurrence rate and metastasis still make the mortality rate of breast cancer high.

[0003] There are many traditional diagnostic methods for breast cancer, among which imaging examinations are more commonly used, such as ultrasound examination, which can observe the internal structure of the breast, and the operation is relatively simple and non-invasive. However, its resolution for some early lesions such as tiny calcifications is limited, and it is easy to miss the diagnosis; X-ray examination (such as molybdenum target examination) can detect some early breast masses and abnormal manifestations such as tiny calcifications, but the detection effect of lesions in dense breast tissue may be affected, and there is a certain radiation hazard. In addition, tissue biopsy is also an important diagnostic method, such as fine needle puncture biopsy, hollow needle puncture biopsy and surgical excision biopsy. Although it can directly obtain lesion tissue for pathological diagnosis and has high accuracy, it is an invasive examination and will bring certain pain to patients. At the same time, its test results also depend on the operator's technical level and experience to a certain extent. If the sampling site is inaccurate, it may lead to misdiagnosis or inability to accurately judge the severity of the disease. Moreover, these traditional diagnostic methods are not ideal for the early detection of tiny lesions as a whole. Abnormalities can often be more clearly found when the tumor develops to a certain stage, which is not conducive to the early and accurate diagnosis of breast cancer.

[0004] At present, the clinical prognosis assessment of breast cancer is mainly based on pathological staging and some protein markers. In terms of pathological staging, factors such as tumor size, lymph node metastasis, and distant metastasis are comprehensively considered to roughly judge the patient's prognosis. However, this method is relatively macroscopic and it is difficult to accurately predict the risk of individual recurrence and metastasis. The subsequent disease progression of some patients in the same stage may vary greatly. At the protein marker level, although markers such as carcinoembryonic antigen (CEA) and carbohydrate antigen 15-3 (CA15-3) can assist in judging the condition, monitoring the treatment effect, and have a certain role in prognosis to a certain extent, their accuracy is limited when used alone, and their accuracy of prognosis is poor. Breast cancer is a highly heterogeneous disease. The existing conventional prognosis assessment methods are difficult to make accurate prognosis judgments for different individuals in a comprehensive and accurate manner, and cannot meet the clinical needs for individualized and accurate prognosis assessment. Therefore, it is very necessary to find more effective new biomarkers to make up for the shortcomings of existing markers. Summary of the invention

[0005] In view of the shortcomings of existing research, the present 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 marker for breast cancer, providing potential targets for the personalized treatment of breast cancer.

[0006] The present invention provides the use of serine palmitoyltransferase small subunit A as a biomarker in preparing a kit for early diagnosis and / or prognosis prediction of breast cancer.

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

[0008] Preferably, the sample comprises 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 in healthy samples is significantly different; the expression level of the serine palmitoyltransferase small subunit A in samples before and after breast cancer treatment intervention is significantly different.

[0010] The present invention also provides the use of a reagent for detecting the small subunit A of serine palmitoyltransferase in preparing a kit for early diagnosis and / or prognosis prediction of breast cancer.

[0011] Preferably, the reagent comprises 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 reagent for detecting the expression level of serine palmitoyltransferase small subunit A comprises an enzyme-linked immunosorbent assay detection reagent and / or a multiplex immunohistochemistry detection reagent.

[0014] Preferably, the ELISA detection reagent and / or the multiplex immunohistochemistry detection reagent comprises an anti-serine palmitoyltransferase small subunit A antibody.

[0015] Beneficial effects:

[0016] The present invention provides the 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. At the same time, the present invention provides the use of a reagent for detecting serine palmitoyltransferase small subunit A in the preparation of a kit for early diagnosis and / or prognosis prediction of breast cancer. Through the analysis of the Cancer Genome Atlas (TCGA) database, the present invention clarifies that SPTSSA mRNA is highly expressed in breast cancer and is closely related to the clinical characteristics of breast cancer (such as tumor size, lymph node metastasis, TNM staging) and poor prognosis, and can be used as a potential factor for predicting poor prognosis of breast cancer patients. At the same time, the present invention finds that SPTSSA is involved in regulating the immune landscape of the tumor microenvironment, and its expression is related to a variety of immune cells (such as CD4+ T cells). 3+ CD 4+ T cells, CD4 68+ CD 163+ It is related to the expression of PD-1, CTLA-4 and macrophages, suggesting that it may play a role in tumor immune escape. Furthermore, the present invention found that the SPTSSA protein level was elevated in the serum of breast cancer patients, and there was a significant difference between preoperative and postoperative for breast cancer patients, indicating that it has potential application value in monitoring tumor load. Based on the above studies, the present 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, and provides a theoretical basis for the treatment strategy targeting SPTSSA. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required to be used in the embodiments are briefly introduced below.

[0018] Figure 1 Schematic diagram of the research method and technical route in Example 1;

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

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

[0021] Figure 4 The Kaplan-Meier survival curve analysis results of the overall survival of breast cancer patients in the SPTSSA high and low expression groups in Example 1 are shown;

[0022] Figure 5 The results of the expression of SPTSSA protein in serum in Example 1 are shown in Figure 1; (a) is the expression of SPTSSA protein in the serum of healthy people and breast cancer patients before surgery, and (b) is 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 curve for the prediction of SPTSSA protein in the preoperative serum of healthy people and breast cancer patients in Example 2;

[0024] Figure 7 2 is the ROC curve for predicting the SPTSSA protein in the preoperative and postoperative serum of breast cancer patients in Example 2. DETAILED DESCRIPTION

[0025] The present invention provides the 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. The accession number of the serine palmitoyltransferase small subunit A in the UniProt database is Q969W0.

[0026] As an embodiment, the kit realizes early diagnosis of breast cancer and / or prediction of breast cancer prognosis by detecting the expression level of the serine palmitoyltransferase small subunit A in the subject sample. As an embodiment, the sample includes but is not limited to a serum sample. As an embodiment, the reagent for detecting the expression level of the serine palmitoyltransferase small subunit A of the present invention is an immunoassay reagent; as another embodiment, the immunoassay reagent includes an enzyme-linked immunosorbent assay (ELISA) detection reagent and / or a multiplex immunohistochemistry (mIHC) detection reagent; as another embodiment, the immunoassay reagent is an enzyme-linked immunosorbent assay detection reagent.

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

[0028] The present invention also provides the use of a reagent for detecting serine palmitoyltransferase small subunit A in preparing a kit for early diagnosis and / or prognosis prediction of breast cancer. As an embodiment, the kit of the present invention is a kit prepared based on immunoassay technology.

[0029] As one embodiment, the reagent of the present invention includes a reagent for detecting the expression level of serine palmitoyltransferase small subunit A; as another embodiment, the reagent for detecting the expression level of serine palmitoyltransferase small subunit A includes an enzyme-linked immunosorbent assay detection reagent and / or a multiplex immunohistochemistry detection reagent; as another embodiment, the enzyme-linked immunosorbent assay detection reagent and / or the multiplex immunohistochemistry detection reagent includes an anti-serine palmitoyltransferase small subunit A antibody.

[0030] The present 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 marker for breast cancer, providing a potential target for the personalized treatment of breast cancer. That is, the present 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, and provides a theoretical basis for the treatment strategy 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 a kit for early diagnosis and / or prognosis prediction of breast cancer is described in detail below in conjunction with the accompanying drawings and examples, but they should not be construed as limiting the scope of protection of the present invention.

[0032] Example 1

[0033] The research method and technical route diagram in this embodiment are as follows Figure 1 shown.

[0034] 1. Test materials and methods

[0035] (1) Data extraction: Pan-Cancer gene set was obtained from The Cancer Genome Atlas (TCGA) to extract the expression data of ENSG00000165389 (SPTSSA) gene in each sample. The RNA sequencing data was used to compare the expression of SPTSSA mRNA in breast cancer tissues and non-tumor tissues, including 1092 breast cancer samples and 113 non-tumor tissue samples.

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

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

[0038] (3) Antibody sources: anti-SPTSSA monoclonal antibody: the anti-SPTSSA monoclonal antibody S1N1 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] Multiplex immunohistochemistry (mIHC) and scoring:

[0040] 1) Bake breast cancer TMA slices in an oven at 70°C for 60 min;

[0041] 2) Xylene dewaxing (vibration for 6 minutes) - 100% alcohol (5 minutes) - 95% alcohol (5 minutes) - 70% alcohol (5 minutes), hydration and then washing with ultrapure water;

[0042] 3) Fix in 10% formalin solution for 10 min and 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 for repair, cover the lid (to prevent the liquid from evaporating during heating), high heat (100% power) for 2.5 minutes, low heat (20% power) for 15 minutes, take out and open the lid, cool naturally to room temperature, rinse with ultrapure water first, and then rinse with TBST;

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

[0045] 6) Anti-SPTSSA monoclonal antibody was diluted to 1:800 with primary antibody diluent (Multiple Fluorescence Immunohistochemistry Kit) (Beijing Bio-Tech Co., Ltd.) and immersed in the sample area at 4°C overnight;

[0046] 7) After antibody recovery, rinse with TBST for 3×2 min, add secondary antibody enhancement solution (horseradish peroxidase (HRP)-conjugated secondary antibody, multiple 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) 100 μl of 1X dye working solution in the optional kit (diluted 1:100 with signal amplification solution) was immersed in the sample area, incubated at room temperature for 10 min in the dark, and rinsed 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 sealing. After repair, cool to a constant temperature, rinse with TBST and then ultrapure water;

[0049] 10) DAPI working solution (multiple fluorescent immunohistochemistry kit) was used to stain and seal the slides;

[0050] 11) TMA was imaged using the Vectra 3.0 automated quantitative pathology imaging system, which consisted of a tissue chip system (UT06, Unitma, South Korea), an inverted microscope (DMIRB, Leica, Germany), and an automatic microplate reader (SN209941, Bio-Tek, USA). Image analysis and scoring were performed using the InForm (Perkin Elmer 2.6.0) software, which set a threshold for positive or negative cells for each cell, calculated the cell percentage in each area, and gave a score (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. The SPTSSA antibody was diluted 1:3000 and coated on a 96-well plate. After incubation, washing, addition of samples and biotinylated secondary antibody detection, TMB substrate was used for color development, and the 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 clinical pathological parameters between the SPTSSA low expression or no expression group and the high expression group. The Kaplan-Meier method was used to evaluate the overall survival differences in patients with different SPTSSA mRNA expression levels, and a Cox regression model was established to predict the effect 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 at P < 0.05. All tests were two-tailed tests, and the data were expressed as mean, standard deviation or 95% confidence interval. SPSS24.0 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 that the SPTSSA mRNA level in breast cancer tissues was significantly increased compared with non-tumor tissues (P<0.05), indicating that SPTSSA is highly expressed in breast cancer tissues. Figure 2 shown.

[0056] The mIHC quantitative analysis based on tissue chips constructed from 145 breast cancer specimens showed that SPTSSA protein was mainly positively expressed in breast cancer cell tissues. Compared with marginal non-tumor tissues, the expression level in tumor tissues was significantly increased, which was consistent with the mRNA expression pattern in TCGA. The clinical prognosis of breast cancer patients in the high expression group was poor.

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

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

[0059] Table 1 Clinical pathological information of 1145 breast cancer specimens

[0060]

[0061]

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

[0063] Table 2 Analysis results of SPTSSA as an independent factor for survival prediction

[0064]

[0065] It can be seen that the SPTSSA protein level is related to the tumor size (x 2 =8.761, P=0.013), degree of lymph node involvement (x 2 =10.713, P = 0.005) and TNM stage (x 2 =9.818, P = 0.007), 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 analysis showed that SPTSSA was significantly correlated with TNM stage (P=0.001), affected overall survival, and was an independent prognostic factor.

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

[0068] Correlation analysis showed that the expression of SPTSSA protein was significantly correlated with CD68 + CD163 + Macrophages (r = 0.234, P = 0.01) and stromal CD3 + CD4 + There was a significant correlation between the presence of T cells (r=0.200, P=0.018).

[0069] Further analysis of mIHC 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 the SPTSSA level in the tumor area of ​​breast cancer, but not in the stromal area. 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 tumor area of ​​breast cancer, and the right figure shows the correlation between the expression of PD-1 and CTLA-4 and the SPTSSA level in the stromal area 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, the Kaplan-Meier survival curve of the overall survival was analyzed and drawn. The results are as follows: Figure 4 shown.

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

[0072] Based on the results of serum ELISA test and analysis of 60 patients before and after surgical intervention, it was found that the level of SPTSSA protein in the serum of breast cancer patients was significantly increased compared with that of 30 healthy controls (P<0.05). Based on the results of serum ELISA test and analysis 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 shown.

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

[0074] 3. Experimental conclusion

[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 staging, etc.) and poor prognosis. It can be used as a potential factor to predict the poor prognosis of breast cancer patients.

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

[0077] (3) The serum SPTSSA protein level in breast cancer patients was elevated and there were 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 characteristics of SPTSSA provide a theoretical basis for the development of therapeutic strategies targeting SPTSSA.

[0079] Example 2

[0080] Sensitivity and specificity of SPTSSA protein as a diagnostic and prognostic marker for breast cancer

[0081] 1. Subjects

[0082] The subject population in the embodiment consists of 30 healthy physical examination people and 72 breast cancer patients, of which preoperative serum samples of 30 healthy physical examination people and 60 breast cancer patients are used to judge the sensitivity and specificity of SPTSSA protein as a diagnostic marker for breast cancer; the preoperative serum samples and postoperative serum samples of the remaining 12 breast cancer patients are used to judge the sensitivity and specificity of SPTSSA as a prognostic marker. The healthy physical examination population comes from the physical examination center of the Affiliated Hospital of Nantong University. The serum of all breast cancer patients was collected when the patient was diagnosed with breast cancer and had not received any chemoradiotherapy or surgical treatment, and was stored in a -80°C refrigerator.

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

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

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

[0086] (3) Blocking: add 1% BSA 200 μl / well and incubate at 37°C in the dark for 2 h;

[0087] (4) Discard the liquid in the wells, wash with PBST for 3 × 3 min, and drain;

[0088] (5) Sample addition: 100 μl / well, incubate at 37°C 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: dilute HRP-SPTSSA 1:1000, 100 μl / well, incubate at 37°C in the dark for 1 h;

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

[0092] (9) Color development: 1× TMB solution (Beijing Solebow Technology Co., Ltd.), 100 μl / well, incubate at 37°C for 1-30 min;

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

[0094] (11) Thermo Scientific Multiskan Go full-wavelength microplate reader was used to measure absorbance at 450 nm.

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

[0096] GraphPadPrism software (version 10.4.0) was used for ROC curve analysis to evaluate the performance of the prediction model. MedCalc (version 23.0.2) software was used for calculation, and the optimal critical value was determined by maximizing the Youden index (sensitivity + specificity - 1). The sensitivity and specificity were 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 (before and after surgery) 68.52％ 81.48％

[0099] The ROC curve analysis results of SPTSSA protein as a diagnostic and prognostic marker for breast cancer are shown in Figure 2. Figure 6 and Figure 7 shown.

[0100] Depend on Figures 6-7 It can be concluded that: the area under the curve of SPTSSA protein in the comparison of preoperative serum of healthy people and breast cancer patients is 0.9108 (0.8587-0.9629), P value <0.001; the area under the curve of SPTSSA protein in preoperative serum and postoperative serum of breast cancer patients is 0.8014 (0.7204-0.8824), P value <0.001. Significance is defined as P <0.05.

[0101] The optimal cutoff, sensitivity and specificity of SPTSSA protein in predicting breast cancer. When the cutoff value was greater than 1.6188, the area under the ROC curve (AUC) of SPTSSA protein in the preoperative serum of healthy and breast cancer patients was 0.9108 (95% CI: 0.8587-0.9629), the sensitivity was 85.00%, and the specificity was 88.33%, which was statistically significant (P<0.001); when the cutoff value was greater than 1.7546, the AUC of SPTSSA protein in the preoperative serum and postoperative serum of breast cancer patients was 0.8014 (95% CI: 0.7204-0.8824), the sensitivity was 68.52%, and the specificity was 81.48%, which was also statistically significant (P<0.001).

[0102] It can be seen that the ROC curve of SPTSSA protein has good diagnostic value in the comparison of serum levels in healthy people 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 embodiment describes the present invention in detail, it is only a part of the embodiments of the present invention, not all of the embodiments. People can also obtain other embodiments based on this embodiment without creativity, 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 a kit for early diagnosis and / or prognosis prediction of breast cancer.

2. The use according to claim 1, characterized in that: The kit realizes early diagnosis of breast cancer and / or prediction of breast cancer prognosis by detecting the expression level of the serine palmitoyltransferase small subunit A in a subject's sample.

3. The use according to claim 2, characterized in that: The sample comprises a serum sample.

4. The use according to claim 2, characterized in that: The serine palmitoyltransferase small subunit A is highly expressed in breast cancer samples and has a significant difference in expression level compared with healthy samples; the expression level of the serine palmitoyltransferase small subunit A has a significant difference in samples before and after breast cancer treatment intervention.

5. Use of a reagent for detecting serine palmitoyltransferase small subunit A in the preparation of a kit for early diagnosis and / or prognosis prediction of breast cancer.

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

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

8. The use according to claim 6, characterized in that: The reagent for detecting the expression level of serine palmitoyltransferase small subunit A comprises an enzyme-linked immunosorbent assay detection reagent and / or a multiplex immunohistochemistry detection reagent.

9. The use according to claim 8, characterized in that: The enzyme-linked immunosorbent assay detection reagent and / or multiple immunohistochemistry detection reagent comprises an anti-serine palmitoyltransferase small subunit A antibody.

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