CN111679072A - Application of KDM6B protein in breast cancer prognosis evaluation kit and diagnosis kit - Google Patents
Application of KDM6B protein in breast cancer prognosis evaluation kit and diagnosis kit Download PDFInfo
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Abstract
The invention provides application of KDM6B protein in a breast cancer prognosis evaluation kit and a breast cancer diagnosis kit, and belongs to the technical field of biology. The application of the KDM6B protein in a breast cancer prognosis evaluation kit comprises the steps of taking the KDM6B protein as a molecular marker, utilizing a KDM6B monoclonal antibody or a KDM6B polyclonal antibody, combining an experimental reagent, and detecting the relative expression quantity of the KDM6B protein in a breast cancer tissue. The invention has the advantages that the KDM6B protein is used as a molecular marker for detecting the expression level of the KDM6B protein and can be used for guiding the prognosis judgment of breast cancer.
Description
Technical Field
The invention belongs to the technical field of biology, and particularly relates to application of KDM6B protein in a breast cancer prognosis evaluation kit and a breast cancer diagnosis kit.
Background
Breast cancer is the most common malignancy in women, second only to lung cancer, with an estimated annual incidence of over 120 million women diagnosed with breast cancer. The incidence of breast cancer in China has risen to the first place of female malignant tumor, and the mortality rate accounts for the fifth place, and becomes one of the biggest threats to female health. Early symptoms of breast cancer are not obvious, diagnosis is already at middle and late stages, and the curative effect of a plurality of breast cancer patients is not ideal after operation, chemotherapy and radiotherapy. Early diagnosis and early treatment of breast cancer and prognostic assessment are key to improving patient survival and extending patient life.
Breast cancer is a highly heterogeneous malignant tumor, and is mainly divided into four subtypes of Luminal a, Luminal B, Her-2 positive and Triple Negative Breast Cancer (TNBC). The triple negative breast cancer is a breast cancer subtype with negative expression of Estrogen Receptor (ER), progestational hormone receptor (PR) and human epidermal growth receptor 2(HER-2), has high malignancy, strong invasiveness and easy occurrence of early metastasis and chemotherapy tolerance, seriously influences the survival of breast cancer patients, and has the 5-year survival rate of less than 15 percent. Due to the lack of endocrine and HER-2-resistant targeted therapeutic receptors, the treatment means of triple negative breast cancer is limited, the prognosis of patients is relatively poor, and an effective index for evaluating the prognosis is still lacking.
Therefore, there is an urgent need in the art to provide new markers for the diagnosis and treatment of breast cancer, and to find genes and/or proteins for the prognosis of breast cancer (especially triple negative breast cancer), and this aspect of research is of great significance for the clinical treatment of breast cancer and the prevention of tumor recurrence.
The typing of breast cancer is complex, especially triple negative breast cancer lacks effective clinical treatment targets, and the prognosis is very poor, so that it is difficult to find effective indexes to reasonably evaluate the treatment effect and prognosis condition of different types of breast cancer. At present, the effect of KDM6B in breast cancer is rarely known, the expression of KDM6B in different types of breast cancer tissues is not clear, and the application of the expression of KDM6B in the prognosis of patients is not reported.
Disclosure of Invention
The invention aims to provide a novel application of KDM6B protein, in particular to an application of KDM6B protein in a breast cancer prognosis evaluation kit and a breast cancer diagnosis kit.
The first object of the present invention can be achieved by the following technical solutions: the application of the KDM6B protein in a breast cancer prognosis evaluation kit is characterized in that the KDM6B protein is used as a molecular marker, a KDM6B monoclonal antibody or a KDM6B polyclonal antibody is used, and an experimental reagent is combined to detect the relative expression amount of the KDM6B protein in a breast cancer tissue.
Preferably, the test reagent is an immunohistochemical test reagent.
Preferably, the KDM6B polyclonal antibody is prepared by KDM6B protein after rabbit immunization.
Preferably, the immunohistochemical reagent comprises phosphate buffered saline solution, sodium citrate solution, ethanol solution, 3% methanol-H2O2Solution, confining liquid, DAB developing liquid and biotin-labeled goat anti-rabbit IgG.
Preferably, the kit is used for detecting the expression level of KDM6B protein in breast cancer tissues in vitro, and the detection method comprises the following steps:
s01: phosphate buffer salt solution, sodium citrate solution, ethanol solution and 3% methanol-H in the kit are utilized2O2Carrying out immunohistochemical staining on the breast cancer tissue section by using solution, confining liquid, DAB developing liquid and biotin labeled goat anti-rabbit IgG;
s02: collecting the dyed image by using an image analysis system;
s03: scoring the staining results respectively by an immunohistochemical integral scoring method;
s04: and (3) dividing KDM6B protein molecules in the breast cancer tissues into high expression quantity and low expression quantity according to the scores.
The second object of the present invention can be achieved by the following technical solutions: the application of the KDM6B protein in a breast cancer diagnosis kit is characterized in that the KDM6B protein is used as a diagnosis marker in the breast cancer diagnosis kit.
The principle of the invention is as follows: the invention mainly expounds the expression characteristics of KDM6B in different types of breast cancer tissues, the relation between the expression of KDM6B and clinical pathological factors, and focuses on the relevance between KDM6B and patient prognosis. The invention provides a new application of KDM6B protein, adopts an immunohistochemical method to detect the expression of KDM6B protein in breast cancer tissues, analyzes the correlation between the expression of KDM6B and breast cancer clinical pathological indexes, and detects the expression quantity of KDM6B protein as a molecular marker based on the correlation between the relative expression quantity and the breast cancer, so as to guide the prognosis of the breast cancer, thereby completing the invention.
Compared with the prior art, the invention has the following advantages:
(1) the invention discloses a protein KDM6B with high expression in breast cancer cells, wherein the significant increase of the expression of KDM6B prompts the existence of breast cancer, and provides a new marker for the diagnosis of breast cancer.
(2) The invention proves that the correlation exists between the KDM6B protein and breast cancer clinical pathological factors through retrospective analysis, and the KDM6B protein is closely related to the occurrence and development of the breast cancer.
(3) According to the invention, the survival curve analysis shows that the different expression levels of KDM6B in triple-negative breast cancer and non-triple-negative breast cancer have different influences on the postoperative survival period of a patient, and the three trends are opposite, wherein in the triple-negative breast cancer, the prognosis of a KDM6B low-expression person is good, but in the non-triple-negative breast cancer, the prognosis of a KDM6B high-expression person is relatively good.
(4) The invention further proves that KDM6B has different prognosis evaluation effects on patients with triple negative breast cancer and non-triple negative breast cancer through the verification of an OSbrca database.
Drawings
FIG. 1 shows that KDM6B protein positive expression site in breast cancer of the present invention is tan (200X);
FIG. 2 shows KDM6B expression in breast cancer cell plasma (400X) according to the invention;
FIG. 3 is KDM6B negative expression (200X) in breast cancer of the invention;
FIG. 4 is KDM6B negative expression (200X) in a breast benign tumor of the present invention;
FIG. 5 is a graph of KDM6B expression of the invention plotted against breast cancer patient survival time (P > 0.05);
FIG. 6 is a graph of KDM6B expression of the invention plotted against the survival time of a patient with triple negative breast cancer (P > 0.05);
FIG. 7 is a plot of KDM6B expression versus survival time for patients with non-triple negative breast cancer according to the invention (P > 0.05);
fig. 8 is a triple negative breast cancer with KDM6B "25% higher" expressing shorter overall survival for patients with TCGA (OS, HR (95% CI) ═ 8.6175(1.0024-74.0813), P ═ 0.0497);
fig. 9 is a graph of the overall survival of KDM6B "25% higher" expressing patients with positive ER expression in non-triple negative breast cancer, GSE7390(OS, HR (95% CI) ═ 0.3164(0.1205-0.8308), P ═ 0.0195);
fig. 10 shows that in non-triple negative breast cancer with positive ER and PR expression, KDM6B "25% higher" expressed patients had longer disease-free survival, GSE21653(DFS, HR (95% CI) ═ 0.3496(0.1341-0.9113), P ═ 0.0315).
Detailed Description
The following are specific embodiments of the present invention and are further described with reference to the drawings, but the present invention is not limited to these embodiments.
Clinical case information collection
Between 2011 and 2014, all 160 patients had histologically confirmed breast cancer and the corresponding molecular subtype from surgical specimens at the second subsidiary hospital of the university of medical science, wenzhou. Recording clinical information of the patient: age, sex, last detailed hospitalization information, tumor size, tumor type and grade, number of lymph node metastases and follow-up information (patient survival, date and cause of death, etc.). Our study was approved by the institutional ethics committee, all patients received written informed consent and authorized surgical removal of tissue for scientific purposes.
Clinical tissue samples
The tumor tissue of the primary breast cancer patient is obtained from the solid part or the hard part of the tumor, and necrotic parts are avoided as much as possible. All patients do not receive any anti-tumor treatment such as radiotherapy or chemotherapy before operation. The histological grade was determined by two independent pathologists, and the clinical staging of tumors was determined according to a clinical classification system. 160 case samples of the study consisted of 80 triple negative breast cancers, 70 non-triple negative breast cancers and 10 benign tissues.
Preparation of reagents
(1) Phosphate buffered saline (0.01mol/L PBS, pH 7.4): 2L of PBS solid powder is slowly dissolved in 1800mL of double distilled water, the pH value of the solution is adjusted to 7.4 by HCl, and then water is added to the solution to reach 2L, and the solution is stored for use at room temperature.
(2) Sodium citrate solution: dissolving sodium citrate buffer solution (0.01mol/L, pH 6.0) in 800mL double distilled water, adjusting the pH value to 6.0, adding water to a constant volume of 1L, and storing at room temperature for use.
(3) Preparing ethanol with different concentrations:
95% ethanol: the preparation proportion is absolute ethanol volume: distilled water volume 19: 1, 475mL of absolute ethanol to 25mL of distilled water, mixing them uniformly and storing at room temperature.
90% ethanol: the preparation proportion is absolute ethanol volume: distilled water volume 9: 1, namely, adding 50mL of distilled water into 450mL of absolute ethyl alcohol, mixing uniformly, and storing at room temperature for use.
85% ethanol: the preparation proportion is absolute ethanol volume: distilled water volume 17: 3, namely 425mL of absolute ethyl alcohol, and 75mL of distilled water are added, mixed uniformly and stored at room temperature for use.
75% of ethanol: the preparation proportion is absolute ethanol volume: distilled water volume 3: 1, 375mL of absolute ethanol, 125mL of distilled water, mixing uniformly and storing at room temperature for use.
(4) 3% methanol-H2O2Solution (3% H for short)2O2): according to 30% H2O2: methanol: and (5) preparing a solution with the required volume by the ratio of 1:1: 8.
(5) DAB color development liquid: 2 drops of the solution B (DAB concentrated solution) and 1 drop of the solution C (enzyme-labeled goat anti-mouse/rabbit IgG polymer) are added dropwise into 1mL of the solution A (DAB substrate buffer), and the mixture is ready to be prepared.
Tumor tissue wax block section
Placing the selected breast cancer wax block sample on an ice bench, precooling for 45min, slicing the wax block sample by using a manual paraffin slicer, wherein the thickness of each slice is 4 mu m, the whole tissue is ensured in the slice, completely placing the paraffin slice in warm water at about 40 ℃, uniformly heating the paraffin slice in the warm water and flatly unfolding the paraffin slice, and fixing the tissue on a slip-proof glass sheet for dyeing.
Immunohistochemical SP method for detecting expression of KDM6B protein
(1) And (3) baking the anti-shedding slide containing the tissue slices in a constant temperature box at 60 ℃ for 90 min.
(2) Paraffin section dewaxing hydration: soaking in xylene for 2 times each for 10min, soaking in 75%, 85%, 90%, and 95% ethanol for 5min, hydrating, washing with phosphate buffer solution on shaking table for 3 times, each for 3 min.
(3) 3% methanol-H was used at room temperature in a dark environment2O2The solution incubated the whole tissue on the sections for 25min to inactivate endogenous peroxidase in the tissue, and then washed 3 times with phosphate buffered saline solution on a shaker for 3min each.
(4) Antigen heat repair: immersing the tissue slices in the prepared antigen retrieval solution (0.01M citrate buffer solution), heating in a pressure cooker until boiling, heating for 10min after boiling, taking out, placing on ice, cooling to room temperature (about 20min), taking out the tissue slices, and washing with phosphate buffer solution for 3 times (5 min each time) on a shaking bed.
(5) Each tissue section is sealed by dripping about 100 mu L of 10% goat serum, so that the goat serum can completely cover the tissue, the tissue is incubated at room temperature for 25min, then the excess serum is thrown off, and the residual serum around the tissue is wiped off by using absorbent paper.
(6) The rabbit anti-human KDM6B primary antibody solution is diluted by PBS according to the proportion, the titer is 1: 500, about 100. mu.L of diluted primary antibody solution was added dropwise to each section so that the primary antibody solution could completely cover the tissue. Rabbit anti-human KDM6B primary antibody, namely rabbit KDM6B polyclonal antibody (can identify KDM6B protein in mammals such as human, mouse and the like).
(7) Placing the tissue slices in a wet box containing water, keeping the slide flat, preventing the first liquid resistance from sliding off and not completely covering the tissue, and placing the tissue slices in a refrigerator at 4 ℃ for 16-24 hours.
(8) The slide was removed and washed 3 times with 5min each time on a shaker with phosphate buffered saline. Then, 50. mu.L of secondary antibody (biotin-labeled goat anti-rabbit IgG) was added dropwise to the sections, the sections were incubated in a 37 ℃ constant temperature water bath for 30min, and the sections were washed 3 times with phosphate buffered saline on a shaker for 5min each time.
(9) And (3) dropwise adding 100 mu L of ready-prepared DAB liquid into each section, developing for about 2-3 min at room temperature, controlling the developing intensity under the observation of a microscope, flushing the DAB liquid by using double distilled water to stop developing when the ideal dyeing intensity is achieved, and flushing residual DAB liquid on the sections by using tap water.
(10) Hematoxylin counterstain for 1min, and washing with tap water for 15 min.
(11) Dehydrating the tissue slices with 95%, 90%, 85% and 75% alcohol, respectively, soaking in xylene for 2 times, each for 5min, and finally sealing with neutral gum and cover glass.
Immunohistochemical staining result scoring
The staining results were scored separately according to immunohistochemical score (IRS) and KDM6B was expressed in the cytoplasm and nucleus of breast cancer, positive expression was a tan pellet. The scoring criteria for immunohistochemical staining requires consideration of both the intensity of staining in the immunoreaction (SI) and the percentage of positive cells (PP).
SI score 4, score 0: no positive cells were found; 1 minute: the strongest part of the positive staining shows weak positive; and 2, dividing: cells with positive staining were visible; and 3, dividing: strongly positively stained cells were visible.
PP was rated 5, with the following scoring criteria, 0: the staining area is not positive; 1 minute: the area of the positive staining area accounts for less than or equal to 25 percent of the area of the whole tissue; and 2, dividing: the area of the positive staining area accounts for 26-50% of the total tissue area; and 3, dividing: the area of the positive staining area accounts for 51-75% of the total tissue area; and 4, dividing: positively stained area > 75%.
The immunohistochemical score is equal to the SI multiplied by the PP, and the results are divided into high expression groups (score is greater than 4 points) and low expression groups (score is more than or equal to 0 and less than or equal to 4 points) according to the score.
Image acquisition and analysis
Immunohistochemical stain images were collected by a Leica-Q550CW image analysis system.
Statistical analysis
Selecting χ according to the counting data2The Kaplan-Meier (KM) method measures survival curves and compares differences in survival rates between groups using the Log-rank test. By single and multifactor analysis (Cox proportional wind)Risk regression model) evaluation of prognostic influencing factors, data analysis using SPSS19.0 software, P<0.05 had a statistical difference.
Breast cancer database analysis
The Gene Expression profile and clinical data in The OSbrca breast Cancer Gene database are stored and managed by The microsoft SQL Server database, The breast Cancer Gene Expression profile database mainly consists of The Cancer Genome Atlas (TCGA) and a Gene Expression integration database (Gene Expression Omnibus, GEO), and The data inclusion analysis is based on The following four criteria: (1) the cohort must have at least 50 breast cancer cases. (2) The cohort must contain individual clinical follow-up information. (3) Probe annotation should be done or the probe can be converted to gene symbol by ID conversion. (4) If the queue has multiple platforms, only platforms with more than 50 individual samples are selected. Survival analysis is carried out on a biomarker KDM6B reported in OSbrca, data are divided into expression of 25% higher VS and 25% lower, the data are divided into two groups of triple negative breast cancer and non-triple negative breast cancer according to the expression conditions of ER, PR and Her-2, a Kaplan-Meier survival curve with a logarithmic rank P value is generated respectively, and HR and a 95% confidence interval (95% CI) are calculated by using single-factor Cox regression analysis.
Expression of KDM6B in breast cancer tissue
Immunohistochemical results show that KDM6B protein is expressed in both cytoplasm and nucleus of breast cancer (as shown in figures 1-4), the high expression cases of KDM6B in 150 breast cancer tissues are 79, the high expression cases of KDM6B in 10 breast benign tissues are 1 and chi-square (chi-square)2) Analysis showed that there was a statistical significance of the difference between the two expressions (P)<0.05), suggesting that KDM6B is expressed in breast cancer tissues more highly than in benign tumors, and the results are shown in table 1.
TABLE 1 differences in KDM6B expression between benign tumors of the breast and breast cancers
Further analysis of the positive expression result of KDM6B showed that 23 of 80 triple negative breast cancers had high expression and 56 of 70 non-triple negative breast cancers had high expression, and the chi-square analysis of the expression difference of KDM6B among breast benign tissue, triple negative breast cancer and non-triple negative breast cancer showed that the difference was statistically significant (P <0.05), suggesting that KDM6B has higher expression in non-triple negative breast cancer tissue compared to triple negative breast cancer, and the results are shown in table 2.
TABLE 2 differences in KDM6B expression between benign breast tumors, non-triple negative and triple negative breast cancers
Relation between KDM6B expression and breast cancer related clinical pathological factors
Analysis of statistical results according to chi-square test shows that in 150 breast cancer patient tissue samples, the expression of KDM6B protein is correlated with the expression of ER, PR, Her-2 and Ki-67 in the breast cancer patient tissues, and the difference has statistical significance (P <0.05) (shown in Table 3); but has no related statistical significance to the diseased age, the tumor diameter and the lymph node metastasis condition of the patient (P > 0.05).
TABLE 3 relationship between KDM6B expression and the clinical pathology factors associated with breast cancer patients
Relation between expression of KDM6B and prognosis of breast cancer patients
To discuss the relationship between the expression of KDM6B and the prognosis of breast cancer patients, based on the results of immunohistochemical staining of KDM6B in breast cancer expression, the results of Kaplan-Meier survival analysis showed that, among the relevant clinical pathological factors of 150 breast cancer patients, the patients' disease age, lymph node metastasis, PR expression and the prognosis of breast cancer patients had a correlation (P <0.05) (as shown in Table 5), and the tumor diameters, ER, Her-2, Ki-67 and KDM6B expression had no statistical correlation (P >0.05) with the prognosis of breast cancer patients. Cox regression model analysis was constructed to assess the correlation between KDM6B expression and age, tumor diameter, lymph node metastasis, ER, PR, Her-2, Ki-67, and the results of Cox model analysis showed that lymph node metastasis is an independent factor affecting the prognosis of patients after breast cancer surgery (as shown in Table 6).
TABLE 5150 relationship between clinical pathological factors and prognosis for breast cancer patients
TABLE 6150 multifactorial analysis of prognosis of breast cancer patients
Relation between KDM6B expression and overall survival time of breast cancer
According to the immunohistochemical staining result of KDM6B expression in the breast cancer and the follow-up condition of the breast cancer patient, the expression of KDM6B in the breast cancer and the condition of the breast cancer patient prognosis are known, and the total survival curve is drawn. Analysis of the curve results showed that there was a difference in post-operative survival for patients with different levels of KEM6B expression in all breast cancer patients, but the difference in survival between the high and low expression groups was not statistically significant (P >0.05) (as shown in figure 5). KDM6B expression in triple negative and non-triple negative breast cancers was further analyzed for prognosis of the breast cancer patient and a total survival curve was plotted. Analysis of curve results shows that patients with different levels of KDM6B expression have different post-operative survival, and KDM6B low-expressor has a better prognosis for higher expressor in triple-negative breast cancer, while KDM6B high-expressor has a relatively better prognosis in later follow-up period, but the survival time of high-expression group and low-expression group is not statistically different (P >0.05) in triple-negative and non-triple-negative breast cancer (as shown in fig. 6 and 7). The survival period is long, and the prognosis effect is good.
Breast cancer database alignment analysis KDM6B relation to breast cancer patient prognosis
The prognostic value of KDM6B gene in breast cancer was analyzed in OSbrca database using existing high throughput data. The results show that in triple negative breast cancer in which ER, PR and Her-2 are all negative, the patients with KDM6B "25% higher" expression had a worse prognosis than "25% lower" expression, Overall Survival (OS) was shorter, and the difference was statistically significant: TCGA (OS, HR (95% CI) ═ 8.6175(1.0024-74.0813), P ═ 0.0497) (as shown in fig. 8), in non-triple negative breast cancers positive for ER expression, KDM6B "higher 25%" expressed patients with longer overall survival than "lower 25%" expression, the difference being statistically significant: GSE7390(OS, HR (95% CI) ═ 0.3164(0.1205-0.8308), P ═ 0.0195) (as shown in fig. 9); in non-triple negative breast cancer with positive ER and PR expression, the patient prognosis of KDM6B expression is better than that of KDM expression of: GSE21653(DFS, HR (95% CI) ═ 0.3496(0.1341-0.9113), P ═ 0.0315) (as shown in fig. 10). The OSbrca database verification analysis shows that KDM6B has different estimated values in different types of breast cancers, the high expression of KDM6B in triple-negative breast cancers indicates poor prognosis, while the high expression of KDM6B in non-triple-negative breast cancers indicates longer survival time, and the difference has statistical significance and indicates that the functional action of KDM6B is possibly related to the expression conditions of ER, PR and Her-2.
The specific embodiments described herein are merely illustrative of the spirit of the invention. Various modifications or additions may be made to the described embodiments or alternatives may be employed by those skilled in the art without departing from the spirit or ambit of the invention as defined in the appended claims.
Claims (6)
- The application of the KDM6B protein in a breast cancer prognosis evaluation kit is characterized in that the KDM6B protein is used as a molecular marker, and a KDM6B monoclonal antibody or a KDM6B polyclonal antibody is combined with an experimental reagent to detect the relative expression amount of the KDM6B protein in breast cancer tissues.
- 2. The use of claim 1, wherein the test agent is an immunohistochemical test agent.
- 3. The use of claim 1, wherein said KDM6B polyclonal antibody is prepared from KDM6B protein by rabbit immunization.
- 4. The use of claim 1, wherein the immunohistochemical assay reagent comprises phosphate buffered saline, sodium citrate solution, ethanol solution, 3% methanol-H2O2Solution, confining liquid, DAB developing liquid and biotin-labeled goat anti-rabbit IgG.
- 5. The use according to claim 1, wherein the kit is used for detecting the expression level of KDM6B protein in breast cancer tissues in vitro, and the detection method comprises the following steps:s01: phosphate buffer salt solution, sodium citrate solution, ethanol solution and 3% methanol-H in the kit are utilized2O2Carrying out immunohistochemical staining on the breast cancer tissue section by using solution, confining liquid, DAB developing liquid and biotin labeled goat anti-rabbit IgG;s02: collecting the dyed image by using an image analysis system;s03: scoring the staining results respectively by an immunohistochemical integral scoring method;s04: and (3) dividing KDM6B protein molecules in the breast cancer tissues into high expression quantity and low expression quantity according to the scores.
- The application of the KDM6B protein in a breast cancer diagnosis kit is characterized in that the KDM6B protein is used as a diagnosis marker in the breast cancer diagnosis kit.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN113943803A (en) * | 2021-10-13 | 2022-01-18 | 深圳市人民医院 | Application of HTR6 in diagnosis and prognosis of breast cancer |
| CN114034866A (en) * | 2021-11-29 | 2022-02-11 | 湖州市中心医院 | Breast cancer diagnosis marker and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN113943803A (en) * | 2021-10-13 | 2022-01-18 | 深圳市人民医院 | Application of HTR6 in diagnosis and prognosis of breast cancer |
| CN114034866A (en) * | 2021-11-29 | 2022-02-11 | 湖州市中心医院 | Breast cancer diagnosis marker and application thereof |
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| CN116312785A (en) * | 2023-01-19 | 2023-06-23 | 首都医科大学附属北京胸科医院 | Breast cancer diagnosis marker gene and screening method thereof |
| CN116539885A (en) * | 2023-07-06 | 2023-08-04 | 上海秤信生物科技有限公司 | Tumor autoantigen/antibody combination for early detection of breast cancer and application thereof |
| CN116539885B (en) * | 2023-07-06 | 2023-09-29 | 上海秤信生物科技有限公司 | Tumor autoantigen/antibody combination for early detection of breast cancer and application thereof |
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