CN111965354A - Application of HO-1 protein in breast cancer prognosis evaluation kit and diagnosis kit - Google Patents
Application of HO-1 protein in breast cancer prognosis evaluation kit and diagnosis kit Download PDFInfo
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- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
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Abstract
The invention provides an application of HO-1 protein in a breast cancer prognosis evaluation kit and a diagnosis kit, belongs to the technical field of biology, and solves the problems that the expression of the existing HO-1 in breast cancer and the effect of nucleus staining in the occurrence and development of the breast cancer are not reported in detail and the like. The invention has the advantages of close relationship between HO-1 protein and the occurrence and development of breast cancer and the like.
Description
Technical Field
The invention belongs to the technical field of biology, and particularly relates to application of HO-1 protein in a breast cancer prognosis evaluation kit and a breast cancer diagnosis kit.
Background
Breast cancer has great heterogeneity and is difficult to diagnose and treat accurately. Breast cancer patients have different Receptor expression profiles, and the most clinically used ones include Estrogen Receptor (ER), Progestin Receptor (PR), and Human Epidermal Growth Factor Receptor 2(Human Epidermal Growth Factor Receptor-2, Her-2). Her-2, which is a tyrosine kinase receptor belonging to the EGFR/ErbB family, is often found to be up-regulated in breast cancer. The Her-2 gene has the greatest correlation with breast cancer, and determines the molecular type classification of the breast cancer together with ER and PR, the breast cancer which is negative to ER, PR and Her-2 is defined as triple negative breast cancer, at least one positive breast cancer is collectively called non-triple negative breast cancer, and the breast cancer can be further classified into Lumina A type, Lumina B type and Her-2 overexpression type according to the expression condition. Molecular typing of breast cancer plays an extremely important role in prognostic evaluation, targeted therapy and the like.
Because the treatment means of breast cancer (especially triple negative breast cancer) is limited, the prognosis of patients is relatively poor, and effective indexes and tools for evaluating the prognosis are not available at present, and in order to realize personalized or more accurate treatment, a more detailed tumor molecular characteristic system needs to be established. The improvement of breast cancer prognosis detection means and the development of a kit can fill such a gap. 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, which is of great significance for the prognosis evaluation of clinical breast cancer.
Although a large number of new anticancer agents are being developed and clinically tested, the treatment of breast cancer is still limited to the targets of ER, PR, Her-2, etc. For example, endocrine therapy is used for ER, PR positive patients, trastuzumab is used for Her-2 high-expression patients, and the like. Therefore, in order to make a breakthrough in the diagnosis, treatment and prognosis evaluation of breast cancer, the search for new markers and targets is of great clinical significance.
Heme Oxidase (HO) plays a crucial role in Heme metabolism, and can decompose and convert Heme into biliverdin, carbon monoxide and ferrous ions, and is a rate-limiting enzyme in Heme metabolism. HO-1 is a member of the stress response enzyme system, and the over-expression of HO-1 exists in tissues and organs under stress (such as inflammation, apoptosis, ischemia and hypoxia), which is an important protection mechanism of cells. In tumor cells, several studies suggest that increased expression plays a role in cancer progression. HO-1 has anti-apoptosis effect in tumor cells, is beneficial to the growth of tumor cells, and plays an important role in malignant transformation of cancer cells. Meanwhile, HO-1 is associated with the development of resistance of tumors to chemotherapy, radiotherapy and photodynamic therapy. Inhibition of HO-1 can restore cancer cells to sensitivity to the above-described treatments.
HO-1 is an endoplasmic reticulum-anchored protein, expressed in the cytoplasm in most cases, but can also be transferred into the nucleus in specific cases. It has now been found that the phenomenon of HO-1 nuclear localization is also seen in prostate, lung and oral cancer cells and is associated with tumor progression. Research of HO-1 in breast cancer is also reported, Rui Deng et al find that HO-1 inhibitor ZnPPIX has the effect of inhibiting the tumor growth of mice in mice inoculated with breast cancer cell strain 4T1, and has the trend of HO-1 expression reduction, which indicates that HO-1 is related to the tumor growth. However, the expression of HO-1 in breast cancer and the role of nuclear staining in the occurrence and development of breast cancer have not been reported in detail.
Disclosure of Invention
The invention aims to solve the problems in the prior art and provides application of HO-1 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 HO-1 protein in the breast cancer prognosis evaluation kit is characterized in that the HO-1 protein is used as a molecular marker, and the relative expression quantity of the HO-1 protein in breast cancer tissues is detected by using a HO-1 monoclonal antibody or a HO-1 polyclonal antibody and combining with an experimental reagent.
Preferably, the test reagent is an immunohistochemical test reagent.
Preferably, the immunohistochemical experimental reagent comprises phosphate buffered saline solution, citrate buffer solution, absolute ethyl alcohol, absolute methyl alcohol, goat serum, DAB developing solution and biotin-labeled goat anti-mouse IgG.
Preferably, the kit is used for detecting the expression amount of the HO-1 protein in breast cancer tissues in vitro, and the detection method comprises the following steps:
s01: carrying out immunohistochemical staining on the breast cancer tissue section by using phosphate buffered saline solution, citrate buffer solution, absolute ethyl alcohol, absolute methyl alcohol, goat serum, DAB color development solution and biotin-labeled goat anti-mouse IgG in the kit;
s02: observing the dyeing result under a bidirectional microscope;
s03: scoring the staining results respectively by an immunohistochemical integral scoring method;
s04: dividing the HO-1 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 HO-1 protein in a breast cancer diagnosis kit is characterized in that the breast cancer diagnosis kit takes the HO-1 protein as a diagnosis marker.
The principle of the invention is as follows: the main content of the invention is to provide a new application of HO-1 protein, an immunohistochemical method is adopted to detect the expression of the HO-1 protein in breast cancer tissues, the correlation between the expression of HO-1 and breast cancer clinical pathological indexes is analyzed, and the detection of the expression quantity of the HO-1 protein serving as a molecular marker can be used for guiding the prognosis judgment of breast cancer based on the correlation between the relative expression quantity of the HO-1 protein and the breast cancer.
Compared with the prior art, the invention has the following advantages:
1. the invention discloses a protein HO-1 highly expressed in breast cancer cells (and the expression is lower in benign tissues), and the significant increase of the HO-1 expression indicates the existence of breast cancer, thereby providing a new marker for the diagnosis of breast cancer. Compared with non-triple negative breast cancer, HO-1 has higher expression in triple negative breast cancer, and the expression is related to breast tumor molecular typing.
2. The invention proves that correlation exists between HO-1 protein and breast cancer clinical pathological factors through retrospective analysis, and suggests that the HO-1 protein is closely related to the occurrence and development of breast cancer.
3. The invention evaluates the staining condition of HO-1 at the tumor subcellular level, and finds that nuclear staining also exists in a few breast cancer specimens. The results of the analysis of variance and comparison with clinical and pathological factors of patients are found to be related to the histological grading of tumors, and HO-1 staining of patients with three grades of histological grading is obviously lower than that of patients with two grades. On molecular typing, the HO-1 cell nucleus staining rate of non-triple negative breast cancer is higher than that of triple negative breast cancer. It is suggested that nuclear localization of HO-1 in breast cancer tends to reduce malignancy of tumors, as a protective mechanism to improve patient prognosis.
4. The invention further proves that the HO-1 expression quantity presents positive correlation with poor prognosis through the verification of an OSbrca database, including overall survival period and disease-free survival period.
5. The invention adopts immunohistochemical staining technology to detect the condition of HO-1 expression in clinical breast cancer and fibroadenoma pathological tissues including cytoplasm and nucleus, retrospectively analyzes the relation between HO-1 and breast cancer clinical pathological characteristics and prognosis, develops the application of HO-1 protein in a breast cancer prognosis evaluation kit, and provides a research basis for future research of HO-1 molecular mechanism in breast cancer and further guidance of clinical diagnosis and treatment.
Drawings
FIG. 1 is a microscope photograph showing the negative staining of a fibroadenoma according to the present invention;
FIG. 2 is a microscope photograph of the present invention showing positive staining of fibroadenoma;
FIG. 3 is a microscope photograph of the invention showing negative staining for non-triple negative breast cancer;
FIG. 4 is a microscope photograph of the invention staining positive for non-triple negative breast cancer;
FIG. 5 is a microscope photograph showing negative staining of triple negative breast cancer according to the present invention;
FIG. 6 is a microscope photograph of the present invention showing positive staining for triple negative breast cancer;
FIG. 7 is a microscope photograph of the invention showing a negative result of a non-triple negative breast cancer nuclear stain;
FIG. 8 is an enlarged view of FIG. 7 of the present invention;
FIG. 9 is a microscope photograph of a non-triple negative breast cancer of the present invention staining positive for nuclei;
FIG. 10 is an enlarged view of FIG. 9 of the present invention;
FIG. 11 is a microscope photograph showing that triple negative breast cancer nuclear staining according to the present invention is negative;
FIG. 12 is an enlarged view of FIG. 11 of the present invention;
FIG. 13 is a microscope photograph of the triple negative breast cancer of the present invention staining positive for nuclear;
FIG. 14 is an enlarged view of FIG. 13 of the present invention;
FIG. 15 is a graph comparing the prognostic evaluation of the 25% of patients with the highest HO-1 expression with the lowest 25% of patients with all types of breast cancer, for a total of 172 cases according to the present invention;
FIG. 16 is a graph comparing the poor prognosis evaluation of 25% of patients with the highest HO-1 expression with the remaining 75% of patients in the data of disease-free survival of breast cancer patients according to the present invention.
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
The method is characterized in that breast cancer modified radical surgery specimens are collected from 2012 to 2016, 139 cases of breast cancer patients are clearly diagnosed in a postoperative pathology department, 10 cases of breast lump excision specimens are clearly diagnosed in 2012, the follow-up time of each patient is 10 months from postoperative to 2019, the longest follow-up time is 93 months, and the shortest follow-up time is 7 months.
Pathological tissue specimen
Pathological sections of 139 breast cancers and 10 breast fibroadenomas of the study were taken from paraffin specimens from 2012 to 2016 in pathology department of the second subsidiary hospital of the university of medical science, wenzhou. The group cases all satisfy the following conditions: (1) the patients in the group all receive breast cancer improvement radical operation; (2) all specimens are subjected to HE staining, and professional pathologists can pathologically confirm breast cancer and fibroadenoma under a light microscope; (3) the examination of all cases exceeds 8 lymph nodes for pathological diagnosis after operation; (4) radiotherapy and chemotherapy are not received before operation in all cases. The patient contact way is acquired through a hospital inpatient medical record information system, follow-up chemotherapy treatment way, outpatient service reexamination and survival condition after breast cancer improved radical operation of the patient are followed up in a way of electric connection, and basic information of the patient and the detailed information of the sample pathology are recorded in detail through the medical record system. The study protocol was approved by the research ethics committee of the second subsidiary hospital of the university of medical science, wenzhou. All patients received written informed consent and authorized surgical removal of tissue for scientific purposes.
Consumable material and source of experimental reagent
Mouse anti-human HO-1 monoclonal antibody (English Abcam company)
Anticreep slide glass (China fir gold bridge)
Cover glass (China fir gold bridge)
Phosphate buffered saline (Beijing Solaibao)
Citrate buffer (Beijing Solaibao)
Anhydrous ethanol (Shanghai Chinese medicine)
Anhydrous methanol (Shanghai Chinese medicine)
Goat serum (Zhonghua golden bridge)
Biotin-labeled goat anti-mouse IgG (China fir gold bridge)
DAB color developing agent (China fir gold bridge)
Immunohistochemical staining method
(1) Precooling the tissue wax block on an ice bench, cutting the tissue wax block into slices with the thickness of 4 mu m after 1 hour, flattening the slices in warm water at the temperature of 40 ℃, and fixing the slices on an anti-falling glass slide for later use.
(2) Baking at 60 ℃ for 2 hours, dewaxing by dimethylbenzene, soaking in 100%, 95%, 85% and 75% alcohol for 5 minutes in sequence for hydration, washing for 3 times by PBS, incubating for 25min at room temperature in the dark by using 3% hydrogen peroxide to inactivate tissue endogenous peroxidase, and washing for 3 times by PBS.
(3) Soaking the slices in sodium citrate buffer solution, placing in a microwave oven, and performing heat-repairing on the antigen for half an hour at medium and high fire. And after the restoration is finished, placing the steel pipe at room temperature until the steel pipe is naturally cooled to room temperature.
(4) Washing with PBS for 3 times, adding 100 μ L10% goat serum, covering tissue, blocking nonspecific antigen, incubating at room temperature for 20min, removing serum, and washing with PBS for 3 times.
(5) Dripping primary antibody (mouse anti-human HO-1 monoclonal antibody) in sequence, keeping the refrigerator at 4 ℃ overnight, placing the refrigerator at room temperature for slow rewarming in the next morning, washing by PBS, adding an enhancer (reagent 2 reaction enhancing liquid in a mouse two-step method detection kit, ZSBB-BIO PV-9002, purchased from China fir gold bridge), placing the refrigerator in a 37 ℃ constant temperature water bath box for 20 minutes, taking out, washing by PBS for 3 times, dripping secondary antibody (biotin-labeled goat anti-mouse IgG), placing the refrigerator in the 37 ℃ constant temperature water bath box for 30 minutes, taking out, washing by PBS for 3 times, performing DAB (digital audio broadcasting), washing by tap water, stopping the color development reaction, dripping hematoxylin the water, performing 1 minute of secondary dyeing, soaking and dewatering in 75%, 85%, 95% and 100% alcohol in sequence, fixing in xylene, and then air-drying, and sealing by neutral gum.
Immunohistochemical criteria
HO-1 is mainly localized to the smooth endoplasmic reticulum of the cytoplasm, and is also stained in a few cell nuclei, and positively expressed as brown granular objects. Immunohistochemical staining results uniform results were determined by two pathologists with more than five years of work experience, after discussion under a two-way microscope, for cases with inconsistent diagnosis. HO-1 immunohistochemical staining is divided into products of staining intensity and staining area, the staining intensity is divided into 0-3 points, negative evaluation is 0 point, weak positive evaluation is 1 point, positive evaluation is 2 points, strong positive evaluation is 3 points; the staining area is rated as 0-4, and no obvious positive cells under the microscope are rated as 0; 0-25% rated 1 point; 25-50% rated 2 points; 50-75% rated 3 points; 75% or more was rated 4 points. The product of the two is obtained by total score, the range is 0-12 scores, the low expression is evaluated when the score is less than or equal to 4 scores, and the high expression is evaluated when the score is more than 4 scores. The cell nuclei were stained positively when the cell nuclei exhibited brown particles, and the cell nuclei were stained negatively when the cell nuclei did not exhibit brown particles.
Data statistical method
Data were analyzed using SPSS (version 18.0). The relationship between HO-1 expression and other breast cancer counting data indexes is tested by chi-square method, the relationship between HO-1 expression and breast cancer survival rate is drawn by Kaplan-Meier method, and the P value less than 0.05 is considered as a meaningful statistical standard.
Breast cancer database analysis
Gene Expression profiles and clinical data in The OSbrca breast Cancer Gene database are stored and managed by The microsoft SQL Server database in The united states, a breast Cancer Gene Expression profile dataset is mainly composed of a tumor Gene profile (The Cancer Genome Atlas, TCGA) and a Gene Expression integrated database (Gene Expression Omnibus, GEO), and 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 symbols 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 the biomarker HO-1 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, Kaplan-Meier survival curves with logarithmic rank P values are generated respectively, and HR and 95% confidence interval (95% CI) are calculated by using single-factor Cox regression analysis.
HO-1 expression in benign breast tissue, non-triple negative breast cancer, triple negative breast cancer
Immunohistochemistry results show that HO-1 is mainly expressed in cytoplasm and is cytoplasmic positive, as shown in FIGS. 1-6, the expression of H0-1 in cells, and FIG. 1 is a microscope picture showing that fibroadenoma staining is negative; FIG. 2 is a microscope photograph showing positive staining of fibroadenoma; FIG. 3 is a micrograph of non-triple negative breast cancer staining negative; FIG. 4 is a micrograph of non-triple negative breast cancer staining positive; FIG. 5 is a micrograph of triple negative breast cancer staining negative; FIG. 6 is a micrograph of triple negative breast cancer staining positive. In the tissue of a patient with fibroadenoma, HO-1 is highly expressed in 1 case, the high expression rate of the patient is 10%, the low expression rate is 9 cases, and the low expression rate is 90%. Among 139 tissues of breast cancer patients, the number of high-expression cases was 67, the high-expression rate was 48.2%, and 72 of breast cancer patients with low-level or no-expression HO-1 staining were present, and the low-expression rate was 51.8%. HO-1 expression was elevated in breast cancer tissue compared to fibroadenoma tissue (P ═ 0.022) (table 1). Among them, 64 cases of non-triple negative breast cancer and 23 cases of high expression rate were 35.9% and 41 cases of low expression rate were 64.1%. 75 patients with triple negative breast cancer, 44 patients with high expression rate, 58.7% of high expression rate, 31 patients with low expression rate and 41.3% of low expression rate. Triple negative breast cancer has a higher HO-expression than non-triple negative breast cancer, with statistical significance (P ═ 0.008) (table 1).
TABLE 1 relationship between HO-1 expression and malignancy and tumor (n-149)
Correlation of HO-1 expression with relevant clinical pathological characteristics
To investigate the clinical and pathological significance of HO-1 expression, we analyzed the correlation of HO-1 expression with clinical and pathological data in 139 breast cancers. HO-1 expression in breast cancer tissues was statistically insignificant (P > 0.05) between different ages, tumor sizes, number of lymph node metastases, histological grade, ER, PR and Ki67 (Table 2); at 110 cases Her-2-Or Her-2+In the case of low expression, HO-1 was expressed at a high level of 59 cases, accounting for 53.6%, and at a low level of 51 cases, accounting for 46.4%. In 29 cases of Her-2++And Her-2+++In the case of high expression, HO-1 was expressed at a high level of 8 cases accounting for 27.6%, and at a low level of 21 cases accounting for 72.4%. The above results show that HO-1 expression correlates with Her-2 expression (P ═ 0.013) (table 2).
TABLE 2 relationship of HO-1 expression to clinical pathological factors of breast cancer patients (n ═ 139)
Relationship between HO-1 nuclear staining and pathological factors of breast cancer patients
HO-1 was not significantly expressed in most breast cancer nuclei, but nuclear expression was seen in a few cases, as shown in FIGS. 7-14, which are microscopic images of H0-1 staining in the nuclei. FIG. 7 is a micrograph of a non-triple negative breast cancer which is negative for nuclear staining; FIG. 8 is an enlarged view of FIG. 7; FIG. 9 is a micrograph of a non-triple negative breast cancer nuclear positive; FIG. 10 is an enlarged view of FIG. 9; FIG. 11 is a micrograph of triple negative breast cancer nuclear staining negative; FIG. 12 is an enlarged view of FIG. 11; FIG. 13 is a micrograph of triple negative breast cancer which is positive for nuclear staining; fig. 14 is an enlarged view of fig. 13.
To investigate the clinical significance between HO-1 nuclear staining and pathological factors of breast cancer patients, we statistically analyzed the correlation between the nuclear expression of HO-1 in 139 cases of breast cancer and clinical pathological data. HO-1 expression in breast cancer cell nuclei was not statistically significant (P > 0.05) compared to Her-2, Ki67 (Table 3); some postoperative pathological specimens of patients are intraductal carcinoma, lobular carcinoma, basal cell subtype, medullary carcinoma and the like, have no histological grading information, and are excluded for statistical analysis, so HO-1 nuclear staining and histological grading statistics are included in 118 cases, wherein the number of 2 levels (three levels are total, the number of 2 levels is less than or equal to 2 represents the first level and the second level) is 69, the number of nuclear staining positives is 17, the positive rate is 24.6%, and the number of nuclear staining negatives is 52. In 49 cases > grade 2 (three grades were shared, > grade 2 means three grades), the nuclear staining was positive in 3 cases, and the positive rate was 6.1%. Statistics shows that the nuclear staining positive rate is higher for the patients with lower histological grade between the two, and the statistics significance is obtained (P is 0.008). Of the 139 cases, 64 cases of non-triple negative breast cancer and 14 cases of positive nuclear staining showed a positive rate of 21.9%. 75 patients with triple negative breast cancer, 7 patients with positive nuclear stain, the positive rate is 9.3%. Compared with non-triple-negative breast cancer, the triple-negative breast cancer has lower HO-1 nuclear staining rate and has statistical significance (P is 0.040). 89 cases of ER negative breast cancer, 9 cases of nuclear positive, the positive rate is 10.1%. The ER positive breast cancer patients are 50, the patients with 12 positive nuclear staining patients have a positive rate of 24.0%. Compared with ER negative breast cancer, the HO-1 nuclear staining rate of the ER positive breast cancer is higher, and the statistical significance is achieved (P is 0.028). 93 cases of PR-negative breast cancer, 9 cases of positive nuclear staining, the positive rate is 9.7%. The positive rate of 46 cases of PR positive breast cancer patients and 12 cases of positive nuclear stain patients is 26.1%. Compared with PR negative breast cancer, the HO-1 nuclear staining rate of the PR positive breast cancer is higher, and the statistical significance is achieved (P is 0.011).
TABLE 3 relationship between HO-1 nuclear staining and pathological factors of breast cancer patients (n ═ 139)
Differential HO-1 expression in OSbrca database and prognostic analysis
The prognostic value of the HO-1 gene in breast cancer was analyzed in the OSbrca database using existing high throughput data. The GSE26338_ GPL1390 database results show that of 172 total breast cancers of all types, the 25% patients with the highest HO-1 expression had a worse prognosis than the 25% with the lowest expression, with shorter OS, a Hazard Ratio (HR) of 12.1614 (95% Cl, 1.5191-97.3582), and the difference was statistically significant (P0.0186) (fig. 15). Similarly, 249 breast cancer patient Disease-Free Survival (DFS) data were counted in the GSE4922_ GPL96 database, with 25% of patients with the highest HO-1 expression having worse prognosis than the remaining 75% with a mean risk ratio of 1.5791 (95% Cl, 1.0135-2.4603) and statistically significant difference (P ═ 0.0435) (fig. 16).
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.
Although terms are used more often herein, the possibility of using other terms is not excluded. These terms are used merely to more conveniently describe and explain the nature of the present invention; they are to be construed as being without limitation to any additional limitations that may be imposed by the spirit of the present invention.
Claims (5)
- The application of the HO-1 protein in a breast cancer prognosis evaluation kit is characterized in that the HO-1 protein is used as a molecular marker, and the relative expression amount of the HO-1 protein in a breast cancer tissue is detected by using a HO-1 monoclonal antibody or a HO-1 polyclonal antibody and combining an experimental reagent.
- 2. The use of claim 1, wherein the test agent is an immunohistochemical test agent.
- 3. The use of claim 2, wherein the immunohistochemical assay reagent comprises phosphate buffered saline, citrate buffered saline, absolute ethanol, absolute methanol, goat serum, DAB staining solution, biotin-labeled goat anti-mouse IgG.
- 4. The use according to claim 1, wherein the kit is used for in vitro detection of the expression level of HO-1 protein in breast cancer tissues, and the detection method comprises the following steps:s01: carrying out immunohistochemical staining on the breast cancer tissue section by using phosphate buffered saline solution, citrate buffer solution, absolute ethyl alcohol, absolute methyl alcohol, goat serum, DAB color development solution and biotin-labeled goat anti-mouse IgG in the kit;s02: observing the dyeing result under a bidirectional microscope;s03: scoring the staining results respectively by an immunohistochemical integral scoring method;s04: dividing the HO-1 protein molecules in the breast cancer tissues into high expression quantity and low expression quantity according to the scores.
- The application of the HO-1 protein in a breast cancer diagnosis kit is characterized in that the breast cancer diagnosis kit takes the HO-1 protein as a diagnosis marker.
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| NORBERTO ARIEL GANDINI ET AL.: "Heme Oxygenase-1 Has an Antitumor Role in Breast Cancer", 《ANTIOXIDANTS & REDOX SIGNALING》 * |
| 刘胜春: "HO-1在良、恶性乳腺组织中的表达及其与血管生成的关系", 《内分泌外科》 * |
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