CN112114144A - Application of reagent for detecting APLN protein expression in preparation of kit for prognosis evaluation of low-grade glioma - Google Patents
Application of reagent for detecting APLN protein expression in preparation of kit for prognosis evaluation of low-grade glioma Download PDFInfo
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- CN112114144A CN112114144A CN202010943536.2A CN202010943536A CN112114144A CN 112114144 A CN112114144 A CN 112114144A CN 202010943536 A CN202010943536 A CN 202010943536A CN 112114144 A CN112114144 A CN 112114144A
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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
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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
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6854—Immunoglobulins
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/52—Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis
Abstract
The reagent for detecting the expression of the APLN protein is used for the low-grade glioma prognostic evaluation, and the low-grade glioma diagnosis, treatment or prognostic evaluation can be assisted by testing the expression level of the APLN in a low-grade glioma patient.
Description
Technical Field
The invention relates to the technical field of biology, in particular to application of a reagent for detecting APLN protein expression in preparation of a kit for low-grade glioma prognosis evaluation.
Background
Glial cells are classified into astrocytes, ependymal cells, and oligodendrocytes. Glioma, abbreviated glioma, also known as glioblastoma, is the most common primary central nervous system tumor, accounting for about 50% of all intracranial primary tumors. Glioma is the most common primary malignant brain tumor in adults, and the 5-year survival rate is 20% -30%. According to the histological criteria described by the World Health Organization (WHO) in 2007, gliomas are classified as grade 4, from the lowest grade I to the highest grade IV. Low-grade gliomas (LGG for Low-grade gliomas abbreviation, grade I-II) are considered to be the most common invasive tumors in the brain, with grade I and II belonging to Low-grade gliomas. Glioblastoma (abbreviated GBM, grade iv) is the most aggressive subtype of glioma.
The low-grade glioma is mostly seen in adults and mostly is grade II glioma, and mainly comprises astrocytoma, oligodendroglioma and oligodendroastrocytoma. Although low-grade glioma grows relatively slowly, the glioma tends to dedifferentiate and transform into high-grade glioma, grows invasively, is easy to relapse after operation and is high in grade, and has high disability rate and disease death rate clinically. In addition, some low-grade gliomas are located in important functional areas in the brain, and have high surgical risks, and are easy to appear or aggravate nerve function damage after the operation.
Therefore, a kit for prognosis evaluation of low-grade glioma is urgently needed to guide clinical realization of individualized treatment, and excessive treatment and insufficient treatment are avoided.
Disclosure of Invention
The invention aims to overcome the defects in the prior art and provides an application of a reagent for detecting APLN protein expression in preparing a kit for low-grade glioma prognosis evaluation.
Preferably, the reagent for detecting the expression of the APLN protein is APLN primary antibody.
The kit comprises APLN primary antibody, a hatching secondary antibody and DAB color development liquid.
The invention has the following beneficial effects: in the invention, the high expression of the APLN is found to be negatively related to the prognosis Overall Survival (OS) and relapse-free survival (RFS) of the low-grade glioma patients, and the high expression of the APLN clearly indicates a shorter survival, namely a poorer prognosis, and indicates that the APLN can be used as a biomarker for prognosis evaluation of the low-grade glioma; meanwhile, the high expression of the APLN is found to be in negative correlation with the prognosis of progression-free survival (PFS) of the low-grade glioma patients, and the high expression of the APLN clearly indicates a shorter survival period, namely a poorer prognosis, which indicates that the APLN can be used as a biomarker for prognosis evaluation of the low-grade glioma. The reagent for detecting the expression of the APLN protein is used in a kit for low-grade glioma prognosis evaluation, and can assist the low-grade glioma diagnosis, treatment or prognosis evaluation by testing the expression level of the APLN in a low-grade glioma patient.
Drawings
FIG. 1 is a graph of the relationship of expression of APLN in an OSlgg assay to the prognostic survival of a patient;
FIG. 2 shows the result of immunohistochemical analysis of APLN expression in low-grade gliomas.
Detailed Description
Example 1
Expression of APLN in brain tissue of patients with low-grade glioma
First, collect 41 cases of glioma patients tumor tissue specimens
Pathological results of all patients prove that the tumor tissues of glioma patients are fixed by 4% paraformaldehyde, and are preserved at 4 ℃ to prepare tissue chips.
Dewaxing and hydrating, specifically, placing the tissue chip in a constant temperature incubator at 60 ℃ for 20min, then soaking in xylene for 10min, replacing xylene and then soaking for 10min, sequentially placing absolute ethyl alcohol for soaking for 10min, soaking absolute ethyl alcohol for 10min again, soaking in 95% alcohol for 10min, soaking in 90% alcohol for 10min, soaking in 85% alcohol for 10min, and soaking in 75% alcohol for 10 min. And (3) antigen retrieval, specifically, adding an antigen retrieval solution citrate, and heating for 10min with medium fire.
Second, immunohistochemical staining
Washing with PBS for 5 min/3 times; blocking with 5% bovine serum albumin for 10min, throwing off, adding APLN primary antibody (anti-apelin-36, 1:150 dilution, Phoenix Biotech Co., Ltd., Beijing, China) dropwise, and standing overnight in a refrigerator at 4 deg.C; incubating the secondary antibody (1:200, marked by horseradish peroxidase (HRP), Severe Biotechnology Ltd., Wuhan, China), dripping DAB color developing solution, observing for 7-8min under a mirror, and stopping dyeing with tap water. Gradient dehydration: 75% ethanol 10s → 85% ethanol 10s → 90% ethanol 10s → 95% ethanol 10s → absolute ethanol I10 s → absolute ethanol II 10s → xylene I5 min → xylene II 5 min; and (6) sealing the sheet.
Third, evaluation of staining
The immunohistochemical staining results were independently evaluated by two pathologists blinded to the patient's clinical pathology parameters. The scores were determined to be positive for APLN, based primarily on tumor cell staining intensity and percentage. APLN staining intensity from 0 to 3 points: 0-negative, 1-weak, 2-moderate, 3-strong. Positive staining rate from 0 to 4 points: 1 part (0-25%), 2 parts (26% -50%), 3 parts (51% -75%) and 4 parts (76% -100%). The total score is derived by multiplying the scale and intensity scores.
The results of the assessment of APLN expression in tumor tissue specimens from glioma patients in this example are shown in Table 1.
TABLE 1
FIG. 1 is a graph of the relationship between expression of APLN and patient prognostic survival as analyzed by OSlgg, and it can be seen from FIG. 1 that APLN-highly expressed patients have significantly shorter prognostic overall survival (OS, p ═ 1E-4; HR: 2.0645; 95% CI:1.4256-2.9897), relapse-free survival (RFS, p ═ 5.80E-3; HR: 1.6931; 95% CI: 1.165-2.4605), progression-free survival (PFS, p ═ 9.50E-3; 1.6897; 95% HR: 1.137-2.511), compared to APLN-lowly expressed low-grade glioma patients, i.e., APLN-high expression is significantly indicative of poorer prognosis.
FIG. 2 shows the result of immunohistochemical analysis of APLN expression in low grade gliomas, and it can be seen from FIG. 2 that the expression level of APLN in grade II low grade gliomas is higher than that in grade I.
According to the analysis, the reagent for detecting the expression of the APLN protein is used in the kit for the low-grade glioma prognosis evaluation, and the evaluation of the low-grade glioma diagnosis, treatment or prognosis can be assisted by testing the expression level of the APLN in a low-grade glioma patient.
Claims (3)
1. Application of a reagent for detecting APLN protein expression in preparation of a kit for prognosis evaluation of low-grade glioma.
2. The use of claim 1, wherein: the reagent for detecting the expression of the APLN protein is APLN primary antibody.
3. The use of claim 1, wherein the kit comprises a APLN primary antibody, a hatching secondary antibody and a DAB staining solution.
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| CN202010943536.2A CN112114144B (en) | 2020-09-09 | 2020-09-09 | Application of reagent for detecting APLN protein expression in preparation of kit for prognosis evaluation of low-grade glioma |
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| CN112114144B CN112114144B (en) | 2021-07-27 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116449010A (en) * | 2023-04-12 | 2023-07-18 | 致远医疗投资(广州)有限责任公司 | Use of FAM111B in diagnosis or prognosis of glioma and related computer-readable medium |
Citations (2)
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| WO2015140296A2 (en) * | 2014-03-20 | 2015-09-24 | Centre National De La Recherche Scientifique (Cnrs) | Use of compounds inhibiting apelin / apj / gp130 signaling for treating cancer |
| CN109825585A (en) * | 2019-03-05 | 2019-05-31 | 江苏省肿瘤医院 | A kind of biomarker of nasopharyngeal carcinoma diagnosis and/or prognosis evaluation |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2015140296A2 (en) * | 2014-03-20 | 2015-09-24 | Centre National De La Recherche Scientifique (Cnrs) | Use of compounds inhibiting apelin / apj / gp130 signaling for treating cancer |
| US20170146518A1 (en) * | 2014-03-20 | 2017-05-25 | Centre National De La Recherche Scientifique (Cnrs) | Use of compounds inhibiting apelin / apj / gp130 signaling for treating cancer |
| CN109825585A (en) * | 2019-03-05 | 2019-05-31 | 江苏省肿瘤医院 | A kind of biomarker of nasopharyngeal carcinoma diagnosis and/or prognosis evaluation |
Non-Patent Citations (3)
| Title |
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| VACHHER M 等: "NAMPT, GRN, and SERPINE1 signature as predictor of disease progression and survival in gliomas", 《J. CELL BIOCHEM.》 * |
| 孔娟 等: "利用网络数据库预测脂肪因子APLN、NUCB2、RARRES2在胶质母细胞瘤中的表达", 《山东大学学报(医学版)》 * |
| 李勇等: "Apelin/APJ在小儿神经母细胞瘤中的表达及意义", 《临床小儿外科杂志》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116449010A (en) * | 2023-04-12 | 2023-07-18 | 致远医疗投资(广州)有限责任公司 | Use of FAM111B in diagnosis or prognosis of glioma and related computer-readable medium |
| CN116449010B (en) * | 2023-04-12 | 2024-03-12 | 致远医疗投资(广州)有限责任公司 | Use of FAM111B in diagnosis or prognosis of glioma and related computer-readable medium |
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