CN113156123A - Application of EphA2 gene in preparation of product for treating or diagnosing breast cancer caused by cell apoptosis-related protein - Google Patents
Application of EphA2 gene in preparation of product for treating or diagnosing breast cancer caused by cell apoptosis-related protein Download PDFInfo
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- CN113156123A CN113156123A CN202110479751.6A CN202110479751A CN113156123A CN 113156123 A CN113156123 A CN 113156123A CN 202110479751 A CN202110479751 A CN 202110479751A CN 113156123 A CN113156123 A CN 113156123A
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- breast cancer
- epha2
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- cell apoptosis
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
- G01N33/57407—Specifically defined cancers
- G01N33/57415—Specifically defined cancers of breast
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
- G01N33/57484—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/74—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving hormones or other non-cytokine intercellular protein regulatory factors such as growth factors, including receptors to hormones and growth factors
Abstract
The invention relates to the technical field of biological medicines, and particularly discloses application of an EphA2 gene in preparation of a breast cancer product for treating or diagnosing protein related to cell apoptosis, wherein the level of cell apoptosis in a breast cancer tissue is reduced, and the expression of EphA2 in the breast cancer tissue is increased and is in negative correlation with the cell apoptosis. The EphA2 gene is in negative correlation with the expressions of NLRP3, Caspase1 and IL-1 beta, and the correlation can be utilized to prepare the medicine for treating invasive breast cancer.
Description
Technical Field
The invention relates to the technical field of biological medicines, in particular to application of an EphA2 gene in preparation of a product for treating or diagnosing breast cancer caused by cell apoptosis-related protein.
Background
Breast cancer is one of the most serious malignant tumors in women, and about one eighth of women all suffer from breast cancer globally. Due to the current changes in living standards and the perfection of screening conditions, the incidence of breast cancer tends to increase year by year. Despite significant advances in surgery, hormonal therapy and molecular targeting in the treatment of breast cancer, the therapeutic effect is effectively improved; there are still significant obstacles and challenges in improving efficacy and survival. An increasing number of studies have shown that inflammation is closely related to the occurrence, development and metastasis of tumors, and that during the formation of tumors, their surrounding tissues secrete cytokines, chemokines and transcription factors to form the Tumor Microenvironment (TME), and these inflammation-related components have been shown to be core factors regulating the anti-Tumor signaling pathway. At present, the invasive breast cancer is diagnosed clinically by taking pathological materials and analyzing whether tissues are invasive breast cancer or not by HE staining, the method not only consumes a long time, but also can not obtain pathological tissues to cause misdiagnosis and is difficult to obtain a medicine for treating the breast cancer associated with the protein related to apoptosis.
Disclosure of Invention
In order to solve the technical problems, the invention provides the application of the EphA2 gene in preparing a product for treating or diagnosing breast cancer caused by cell apoptosis-related proteins, wherein the expression level of the EphA2 gene is in negative correlation with the cell apoptosis-related proteins NLRP3, Caspase1 and IL-1 beta.
The invention provides an application of EphA2 gene in preparing a product for treating or diagnosing breast cancer caused by cell apoptosis-related protein.
Further, the breast cancer is invasive breast cancer.
Further, the cell apoptosis-related protein is NLRP 3.
The invention also provides application of an expression product of the EphA2 gene in preparing a medicament for treating breast cancer caused by NLRP 3.
Further, the cell apoptosis-related protein is Caspase 1.
The invention also provides application of an expression product of the EphA2 gene in preparing a medicament for treating breast cancer caused by Caspase 1.
Further, the protein related to apoptosis of cells is IL-1 beta.
The invention also provides application of an expression product of the EphA2 gene in preparing a medicament for treating breast cancer caused by IL-1 beta.
The invention also provides application of the EphA2 gene in preparing a kit for diagnosing breast cancer.
Compared with the prior art, the invention has the beneficial effects that:
1. the invention provides application of EphA2 gene in preparing a medicament for treating breast cancer caused by cell apoptosis-related proteins (NLRP3, Caspase1 and IL-1 beta).
2. The invention provides application of an expression product of EphA2 gene in preparing a medicament for treating breast cancer caused by cell apoptosis-related proteins (NLRP3, Caspase1 and IL-1 beta).
3. The invention provides application of EphA2 gene in preparation of a kit for diagnosing invasive breast cancer.
4. The expression level of the EphA2 gene is in negative correlation with cell apoptosis-related proteins (NLRP3, Caspase1 and IL-1 beta), so that the expression of the cell apoptosis-related proteins (NLRP3, Caspase1 and IL-1 beta) can be increased by reducing the expression of the EphA2 in breast cancer tissues, the cell apoptosis level in the breast cancer tissues is increased, the cancer cell apoptosis is promoted, the proliferation of cancer cells is inhibited, and the effect of treating invasive breast cancer is achieved.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the drawings without creative efforts.
FIG. 1 is a diagram showing the expression of NLRP3 in normal breast tissue, paracancerous tissue and breast cancer tissue according to the present invention, wherein 1 is normal breast tissue, 2 is paracancerous tissue, and 3 is breast cancer tissue;
wherein, FIG. 1a is an expression gel diagram of NLRP3 in normal breast tissue, paracarcinoma tissue and breast cancer tissue;
FIG. 1b is a statistical graph of the expression levels of NLRP3 in normal breast, paracancerous and breast cancer tissues in the present invention;
FIG. 2 is a diagram showing the expression of Caspase1 and IL-1 β in normal breast tissue, paracancerous tissue and breast cancer tissue in the present invention, wherein 1 is normal breast tissue, 2 is paracancerous tissue, and 3 is breast cancer tissue;
wherein, FIG. 2A is a gel diagram of Caspase1 and IL-1 beta expression in normal breast tissue, paracancerous tissue and breast cancer tissue;
FIG. 2B is a statistical graph of the expression levels of Caspase1 and IL-1 β in normal breast, paracancerous and breast cancer tissues;
FIG. 3 is a graph showing the expression of EphA2 in normal breast tissue, paraneoplastic tissue and breast cancer tissue according to the present invention, wherein 1 is normal breast tissue, 2 is paraneoplastic tissue, and 3 is breast cancer tissue;
wherein FIG. 3a is a glue map of EphA2 expression in normal breast tissue, paraneoplastic tissue and breast cancer tissue;
FIG. 3b is a statistical plot of the expression levels of EphA2 in normal breast tissue, paraneoplastic tissue and breast cancer tissue;
FIG. 4 is a graph showing the correlation analysis of EphA2 with NLRP3, Caspase1 and IL-1. beta. for breast cancer patients according to the present invention;
wherein, FIG. 4A shows a correlation analysis of EphA2 with NLRP3 for breast cancer patients;
FIG. 4B shows a correlation analysis of EphA2 with Caspase1 in breast cancer patients;
FIG. 4C shows a correlation analysis of EphA2 with IL-1. beta. for breast cancer patients.
Detailed Description
The following detailed description of specific embodiments of the invention is provided, but it should be understood that the scope of the invention is not limited to the specific embodiments. All other embodiments, which can be obtained by a person skilled in the art without any inventive step based on the embodiments of the present invention, are within the scope of the present invention. The experimental methods described in the examples of the present invention are all conventional methods unless otherwise specified, and materials, reagents and the like used in the following examples are commercially available unless otherwise specified.
Example 1
Correlation analysis of EphA2, NLRP3, Caspase1 and IL-1 beta in various breast cancer tissues
1. Subject: 42 specimens of breast cancer, tissues beside the cancer and normal breast tissue of the general hospital tumor hospital of Ningxia medical university are collected. The pathological type is invasive breast cancer, the experiment is approved by ethical committee of hospital, and informed consent of patients is obtained
2. Instrument materials and reagents
The main apparatus is as follows: clean bench (antai); model 5415D micro bench centrifuge (Eppendorf); precision balance model BS110S (Sartorius); vertical electrophoresis and Model 680 full-automatic enzyme labeling machine (Bio-Rad); homogenizers (MP Biomedicals); laser confocal microscopy (Zeiss);
the main reagents are as follows: protein extraction kit and protein quantification kit (Nanjing, Kaiyi); EphA2 and ICAM1 antibodies (Affinity, Jiangsu); caspase1, NLRP3 and IL-1 β antibody (US, CST); PVDF membranes (Millipore, usa); clean bench (suzhou, anta); homogenizers (MP Biomedicals, usa); model 5415D micro bench centrifuge (Eppendorf, Germany); precision balances of the BS110S type (Sartorius, germany); vertical electrophoresis and Model 680 full-automatic enzyme labeling machine (Bio-Rad, USA).
3. Method of producing a composite material
(1) Western blot detection of protein expression of NLRP3, Caspase1 and IL-1 beta
Extracting the whole protein of each group of mammary tissues according to the specification of the whole protein extraction kit, quantifying by using a BCA method, adding a sample buffer solution, boiling and denaturing for 5 min; taking 40 mu g of total protein from each group, carrying out SDS-PAGE gel electrophoresis for 80V, then electrically transferring to a PVDF membrane, and sealing for 2h by 5% skimmed milk; incubating with anti-NLRP 3, Caspase1 and IL-1 beta antibody at 4 deg.C overnight at a dilution ratio of 1:1000, incubating with secondary antibody at room temperature for 2h at a dilution ratio of 1:5000, and washing with PBST for 3 times, each for 10 min. Scanning by a gel image analysis imaging system, taking beta-actin as an internal reference, and calculating the ratio of the gray values of NLRP3, Caspase1, IL-1 beta and the internal reference of the beta-actin to perform semi-quantitative detection.
(2) Western blot detection of protein expression of EphA2
Extracting the whole protein of each group of mammary tissues according to the specification of the whole protein extraction kit, quantifying by using a BCA method, adding a sample buffer solution, boiling and denaturing for 5 min; taking 40 mu g of total protein from each group, carrying out SDS-PAGE gel electrophoresis for 80V, then electrically transferring to a PVDF membrane, and sealing for 2h by 5% skimmed milk; incubated with anti-EphA 2 antibody at a dilution ratio of 1:1000 overnight at 4 deg.C, incubated with secondary antibody at a dilution ratio of 1:5000 for 2h at room temperature, and washed 3 times with PBST for 10min each. Scanning by a gel image analysis imaging system, taking beta-actin as an internal reference, and calculating the ratio of the EphA2 to the gray value of the beta-actin internal reference for semi-quantitative detection.
(3) Correlation analysis of EphA2 and cell apoptosis-related proteins NLRP3, Caspase1 and IL-1 beta
Correlation analysis was performed on breast cancer tissues using Prism 6.0 statistical software.
(4) Statistical treatment
The experimental results are all measured data, the experimental data are subjected to statistical analysis by using Prism 6.0 statistical software, and the experimental results of the measured data are calculated as mean +/-standard deviation
Showing that the comparison between two groups adopts t test, the comparison between three groups adopts one-factor analysis of variance, P<0.05 indicates that the difference is statistically significant.
4. Results
(1) Expression of apoptosis-related protein NLRP3 in various tissues of breast cancer patients
To demonstrate whether cellular apoptosis plays a role in breast cancer, Western blot was used to test the protein expression level of cellular apoptosis-associated protein NLRP3 in each group. As shown in FIG. 1, the expression of NLRP3(P <0.005) protein in breast cancer tissue is reduced compared with normal breast tissue and para-cancer tissue, and the difference is statistically significant.
(2) Expression of apoptosis-related proteins Caspase1 and IL-1 beta in various tissues of breast cancer patients
Western blot was used to determine the protein expression levels of Caspase1 and IL-1. beta. in each group, and the results are shown in FIG. 2: compared with normal breast tissues and tissues beside the cancer, the expression of Caspase1 protein in the breast cancer tissues is reduced (P <0.005) and the expression of IL-1 beta is reduced (P <0.05, P <0.005), and the difference has statistical significance.
(3) EphA2 expression in various groups of breast cancer patients
The expression of EphA2 protein in each group is detected by Western blot, and the result is shown in FIG. 3: EphA2 protein expression levels were elevated in breast cancer tissues compared to normal breast tissues and paraneoplastic tissues (P <0.01, P < 0.05). The differences are statistically significant.
(4) Correlation analysis of EphA2 protein expression and cell apoptosis-related proteins NLRP3, Caspase1 and IL-1 beta in various groups of breast cancer patients
To verify whether EphA2 is associated with the level of cellular apoptosis in each tissue of breast cancer, we performed pearson correlation analysis on EphA2 protein expression and NLRP3, Caspase1 and IL-1 β apoptosis-related proteins in 45 breast cancer patient tissues, and found that EphA2 is negatively correlated with NLRP3, Caspase1 and IL-1 β (as shown in FIG. 4).
In conclusion, the level of cell apoptosis in the breast cancer tissue is reduced, the expression of EphA2 in the breast cancer tissue is increased, and the expression is in negative correlation with the cell apoptosis, and the level of cell apoptosis in the breast cancer tissue can be increased by regulating the expression of EphA2, so that the effects of promoting apoptosis and inhibiting cancer cell proliferation are achieved on invasive breast cancer cells.
While preferred embodiments of the present invention have been described, additional variations and modifications in those embodiments may occur to those skilled in the art once they learn of the basic inventive concepts. Therefore, it is intended that the appended claims be interpreted as including preferred embodiments and all such alterations and modifications as fall within the scope of the invention.
It will be apparent to those skilled in the art that various changes and modifications may be made in the present invention without departing from the spirit and scope of the invention. Thus, if such modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include such modifications and variations.
Claims (9)
- Use of the EphA2 gene in the preparation of a product for treating or diagnosing breast cancer caused by a protein associated with apoptosis of cells.
- 2. The use of the EphA2 gene of claim 1 for the preparation of a product for the treatment or diagnosis of breast cancer caused by a protein associated with apoptosis in cells, wherein the breast cancer is invasive.
- 3. The use of the EphA2 gene of claim 2 for the preparation of a medicament for the treatment of breast cancer caused by a cell apoptosis-related protein that is NLRP 3.
- 4. Use of the expression product of EphA2 gene of claim 3 in the manufacture of a medicament for treating breast cancer caused by NLRP 3.
- 5. The use of the EphA2 gene of claim 1 for the manufacture of a medicament for the treatment of breast cancer caused by a cellular apoptosis-related protein, wherein the cellular apoptosis-related protein is Caspase 1.
- 6. Use of the expression product of the EphA2 gene of claim 5 in the preparation of a medicament for treating breast cancer caused by Caspase 1.
- 7. The use of the EphA2 gene of claim 1 for the manufacture of a medicament for the treatment of breast cancer caused by a protein associated with cellular apoptosis, wherein the protein associated with cellular apoptosis is IL-1 β.
- 8. Use of the expression product of the EphA2 gene of claim 7 in the manufacture of a medicament for treating breast cancer caused by IL-1 β.
- 9. Use of the EphA2 gene of claim 1 in the preparation of a kit for diagnosing breast cancer.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114622012A (en) * | 2022-04-21 | 2022-06-14 | 宁夏医科大学 | EphA2 gene and application of methylation level detection reagent thereof |
Citations (3)
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| US20130344486A1 (en) * | 2011-02-28 | 2013-12-26 | Rheinische Friedrich-Wilhelms-Universitat Bonn | Method of diagnosing breast carcinoma |
| CN107543926A (en) * | 2017-08-25 | 2018-01-05 | 复旦大学附属金山医院 | A kind of application of new Diagnosis of Breast invasive ductal carcinoma |
| CN110840887A (en) * | 2019-11-18 | 2020-02-28 | 杭州彗搏科技有限公司 | Application of triclabendazole in preparation of medicine for treating breast cancer |
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- 2021-04-30 CN CN202110479751.6A patent/CN113156123B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20130344486A1 (en) * | 2011-02-28 | 2013-12-26 | Rheinische Friedrich-Wilhelms-Universitat Bonn | Method of diagnosing breast carcinoma |
| CN107543926A (en) * | 2017-08-25 | 2018-01-05 | 复旦大学附属金山医院 | A kind of application of new Diagnosis of Breast invasive ductal carcinoma |
| CN110840887A (en) * | 2019-11-18 | 2020-02-28 | 杭州彗搏科技有限公司 | Application of triclabendazole in preparation of medicine for treating breast cancer |
Non-Patent Citations (3)
| Title |
|---|
| LINGZHI ZHOU等: "EphA2 as a new target for breast cancer and its potential clinical application" * |
| M. PAN: "Overexpression of EphA2 gene in invasive human breast cancer and its association with hormone receptor status" * |
| 赵玉斌: "乳腺癌EphA2，EphrinA1，ERα，ERβ和PR的表达及RNAi沉默EphA2基因的实验研究" * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114622012A (en) * | 2022-04-21 | 2022-06-14 | 宁夏医科大学 | EphA2 gene and application of methylation level detection reagent thereof |
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