WO2023217035A1 - 检测c1qbp蛋白表达水平的试剂在制备口腔癌筛查或预后试剂盒中的用途 - Google Patents
检测c1qbp蛋白表达水平的试剂在制备口腔癌筛查或预后试剂盒中的用途 Download PDFInfo
- Publication number
- WO2023217035A1 WO2023217035A1 PCT/CN2023/092512 CN2023092512W WO2023217035A1 WO 2023217035 A1 WO2023217035 A1 WO 2023217035A1 CN 2023092512 W CN2023092512 W CN 2023092512W WO 2023217035 A1 WO2023217035 A1 WO 2023217035A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- c1qbp
- reagent
- expression level
- detecting
- oral
- Prior art date
Links
- 239000003153 chemical reaction reagent Substances 0.000 title claims abstract description 34
- 238000012216 screening Methods 0.000 title claims abstract description 18
- 208000003445 Mouth Neoplasms Diseases 0.000 title claims abstract description 16
- 208000012987 lip and oral cavity carcinoma Diseases 0.000 title claims abstract description 16
- 238000004393 prognosis Methods 0.000 title claims abstract description 16
- 238000002360 preparation method Methods 0.000 title claims abstract description 7
- 102000004169 proteins and genes Human genes 0.000 title description 4
- 108090000623 proteins and genes Proteins 0.000 title description 4
- 102100037078 Complement component 1 Q subcomponent-binding protein, mitochondrial Human genes 0.000 claims abstract description 58
- 101150060120 C1qbp gene Proteins 0.000 claims abstract description 55
- 208000000102 Squamous Cell Carcinoma of Head and Neck Diseases 0.000 claims abstract description 33
- 201000002740 oral squamous cell carcinoma Diseases 0.000 claims abstract description 33
- 210000002200 mouth mucosa Anatomy 0.000 claims abstract description 11
- 238000011532 immunohistochemical staining Methods 0.000 claims description 9
- 238000002965 ELISA Methods 0.000 claims description 4
- 238000012360 testing method Methods 0.000 claims description 2
- 239000003550 marker Substances 0.000 abstract description 3
- 238000000338 in vitro Methods 0.000 abstract description 2
- 210000001519 tissue Anatomy 0.000 description 13
- 238000000034 method Methods 0.000 description 9
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 9
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 8
- 206010028980 Neoplasm Diseases 0.000 description 8
- 210000004027 cell Anatomy 0.000 description 7
- 239000000243 solution Substances 0.000 description 7
- CTQNGGLPUBDAKN-UHFFFAOYSA-N O-Xylene Chemical compound CC1=CC=CC=C1C CTQNGGLPUBDAKN-UHFFFAOYSA-N 0.000 description 6
- 201000011510 cancer Diseases 0.000 description 6
- 239000012188 paraffin wax Substances 0.000 description 6
- 239000008096 xylene Substances 0.000 description 6
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 5
- 230000003211 malignant effect Effects 0.000 description 5
- 210000004400 mucous membrane Anatomy 0.000 description 5
- 239000001993 wax Substances 0.000 description 5
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 4
- 239000000427 antigen Substances 0.000 description 4
- 102000036639 antigens Human genes 0.000 description 4
- 108091007433 antigens Proteins 0.000 description 4
- 201000010099 disease Diseases 0.000 description 4
- 239000012153 distilled water Substances 0.000 description 4
- 230000002503 metabolic effect Effects 0.000 description 4
- 238000001514 detection method Methods 0.000 description 3
- 210000003470 mitochondria Anatomy 0.000 description 3
- 230000008439 repair process Effects 0.000 description 3
- 206010041823 squamous cell carcinoma Diseases 0.000 description 3
- WZUVPPKBWHMQCE-UHFFFAOYSA-N Haematoxylin Chemical compound C12=CC(O)=C(O)C=C2CC2(O)C1C1=CC=C(O)C(O)=C1OC2 WZUVPPKBWHMQCE-UHFFFAOYSA-N 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- 206010027476 Metastases Diseases 0.000 description 2
- 229930040373 Paraformaldehyde Natural products 0.000 description 2
- 230000004791 biological behavior Effects 0.000 description 2
- 230000002596 correlated effect Effects 0.000 description 2
- 230000018044 dehydration Effects 0.000 description 2
- 238000006297 dehydration reaction Methods 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 230000009401 metastasis Effects 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 229920002866 paraformaldehyde Polymers 0.000 description 2
- 230000008672 reprogramming Effects 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 238000010186 staining Methods 0.000 description 2
- 210000004881 tumor cell Anatomy 0.000 description 2
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 description 1
- 206010006187 Breast cancer Diseases 0.000 description 1
- 208000026310 Breast neoplasm Diseases 0.000 description 1
- 206010008342 Cervix carcinoma Diseases 0.000 description 1
- 101000740725 Homo sapiens Complement component 1 Q subcomponent-binding protein, mitochondrial Proteins 0.000 description 1
- 206010058467 Lung neoplasm malignant Diseases 0.000 description 1
- 208000007433 Lymphatic Metastasis Diseases 0.000 description 1
- 208000006105 Uterine Cervical Neoplasms Diseases 0.000 description 1
- 235000011114 ammonium hydroxide Nutrition 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000006399 behavior Effects 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 230000009400 cancer invasion Effects 0.000 description 1
- 201000010881 cervical cancer Diseases 0.000 description 1
- 210000001072 colon Anatomy 0.000 description 1
- 210000000805 cytoplasm Anatomy 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 230000037149 energy metabolism Effects 0.000 description 1
- 230000017188 evasion or tolerance of host immune response Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 210000002216 heart Anatomy 0.000 description 1
- 238000003384 imaging method Methods 0.000 description 1
- 238000003364 immunohistochemistry Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000003902 lesion Effects 0.000 description 1
- 201000005202 lung cancer Diseases 0.000 description 1
- 208000020816 lung neoplasm Diseases 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 230000004066 metabolic change Effects 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 210000001672 ovary Anatomy 0.000 description 1
- 230000010627 oxidative phosphorylation Effects 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 238000001959 radiotherapy Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 210000002027 skeletal muscle Anatomy 0.000 description 1
- 210000000813 small intestine Anatomy 0.000 description 1
- 239000001509 sodium citrate Substances 0.000 description 1
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 1
- 238000007619 statistical method Methods 0.000 description 1
- 238000001356 surgical procedure Methods 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 230000009885 systemic effect Effects 0.000 description 1
- 239000008399 tap water Substances 0.000 description 1
- 235000020679 tap water Nutrition 0.000 description 1
- 210000001550 testis Anatomy 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 230000004614 tumor growth Effects 0.000 description 1
- 239000000439 tumor marker Substances 0.000 description 1
Classifications
-
- 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
- G01N33/6893—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids related to diseases not provided for elsewhere
-
- 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
-
- 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
-
- 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
-
- 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
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
- G01N2333/46—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans from vertebrates
- G01N2333/47—Assays involving proteins of known structure or function as defined in the subgroups
-
- 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
Definitions
- the present invention relates to the field of in vitro diagnostic reagents, and specifically relates to the use of reagents for detecting C1QBP protein expression levels in the preparation of oral cancer screening or prognosis kits.
- OSCC Oral squamous cell carcinoma
- lymph node metastasis the mechanism causing the biological behavior of malignant metastasis in OSCC is still unclear.
- Current research shows that there are complex metabolic changes in tumor cells.
- the metabolic heterogeneity that exists during the development of tumors is tumor metabolic reprogramming.
- Tumor metabolic reprogramming is one of the important hallmarks of malignant tumors and is related to tumor growth, invasion, and metastasis. It is closely related to malignant behaviors such as immune evasion, and metabolism-targeted tumor treatment has huge clinical application potential.
- C1QBP is also known as gClqR, p32, p33 and HABP1.
- C1QBP can be distributed in cells and on the cell surface, and can also be secreted out of cells as a secreted protein. In cells, C1QBP is mainly located in mitochondria. C1QBP in mitochondria Its important function is to maintain oxidative phosphorylation, provide energy to cells and maintain metabolic balance. C1QBP is highly expressed in the mitochondria of a large number of cells that require energy metabolism (such as heart, skeletal muscle, testis, ovary, small intestine and colon). Current research shows that the expression of C1QBP is increased in a variety of malignant tumors, such as breast cancer and lung cancer.
- C1QBP is low-expressed in cervical cancer, indicating that C1QBP regulates tumor biological behavior in a tissue-specific manner.
- C1QBP Studies on expression and function in OSCC have not been reported yet.
- the object of the present invention is to provide a new protein oral cancer marker and the use of a detection reagent for the marker in preparing a kit for oral cancer screening or prognosis.
- C1QBP protein in the preparation of oral cancer screening or prognostic markers.
- the reagent for detecting the expression level of C1QBP protein is a reagent for enzyme-linked immunosorbent assay.
- the reagent for detecting the expression level of C1QBP protein is a reagent for immunohistochemical staining test.
- the oral cancer is oral squamous cell carcinoma.
- the reagent for detecting the expression level of C1QBP protein is a reagent for detecting the expression level of C1QBP protein in human oral mucosa tissue.
- the invention also provides a kit for oral cancer screening or prognosis, which includes a reagent for detecting the expression level of C1QBP protein.
- the reagent for detecting the expression level of C1QBP protein is a reagent for enzyme-linked immunosorbent assay.
- the reagent for detecting the expression level of C1QBP protein is a reagent for immunohistochemical staining.
- the oral cancer is oral squamous cell carcinoma.
- the reagent for detecting the expression level of C1QBP protein is a reagent for detecting the expression level of C1QBP protein in human oral mucosa tissue.
- the key to the present invention is to determine that the expression level of C1QBP in human oral mucosal tissue is significantly related to the risk of oral squamous cell carcinoma. Therefore, the risk of oral squamous cell carcinoma can be determined by detecting the expression level of C1QBP in human oral mucosal tissue. Risk.
- the specific means of detecting the expression level of C1QBP in human blood various means disclosed in the prior art can be used.
- the immunohistochemical staining method is specifically used for detection, but it is not limited to this method. Any method that can detect C1QBP
- the expression level methods can be used for oral squamous cell carcinoma screening or prognosis.
- the present invention provides a new oral squamous cell carcinoma screening marker and a new oral squamous cell carcinoma screening kit, which can realize effective screening of oral squamous cell carcinoma and predict the course and outcome of the disease, and has the characteristics Good application prospects.
- FIG. 1 Protein expression patterns of C1QBP in various stages of oral mucosal cancer (A. Expression of C1QBP in normal oral mucosa, oral potentially malignant diseases (OPMD) and oral squamous cell carcinoma (OSCC); B. C1QBP expression changes with oral mucosa Progress and changes of mucosal cancer)
- normal oral mucosal tissue, oral potentially malignant disease (OPMD) patient tissue, and OSCC patient lesion tissue were collected, including 10 cases in the normal control group, 13 cases in the OPMD group, and 45 cases in the OSCC group.
- the obtained tissues were fixed, dehydrated, and embedded according to the following methods, and paraffin sections were made.
- the obtained tissue sections were subjected to immunohistochemical staining according to the following method.
- Antigen repair Prepare a pressure cooker, add water, and place an antigen repair box containing the antigen repair solution. This experiment used sodium citrate buffer (10mM, pH6.0). If the pot lid is not tightly closed, heat to boiling in hot pot mode. Place the paraffin sections into the antigen retrieval box, close the lid tightly, heat to steam and time for 3 minutes. Slowly deflate until no gas comes out, remove the air valve, and take out the box. Uncover the box and place it under cold water to cool for 20 minutes. TBS rinse, 5 min, 2 times.
- C1QBP takes the presence of yellow or brown particles in the cytoplasm as positive, and evaluates the average staining intensity of the entire section and the proportion of positive cells in tumor cells in each field.
- Immunohistochemistry staining score was performed on the percentage. All sections were blindly tested by two experienced researchers, who evaluated and calculated an average score for each section. The scoring criteria are as follows (Table 1 and Figure 1)
- the expression level of C1QBP in normal oral mucosal tissues was relatively low (average score 0.900)
- the expression level of C1QBP in lesional tissues of patients with oral potential malignant diseases (OPMD) was enhanced (average score 3.538)
- the expression level of C1QBP in OSCC patient tissues was Expression of water is relatively strongest (average score 6.067)
- the OPMD group has statistical significance compared with the normal oral mucosa group (P ⁇ 0.05)
- the OSCC group has statistically significant compared with the normal oral mucosa group (P ⁇ 0.001)
- the OSCC group was statistically significant compared with the OPMD group (P ⁇ 0.01)
- the expression level of C1QBP was negatively correlated with the prognosis of OSCC patients, and the differences were statistically significant (P ⁇ 0.05).
- Example 2 The composition of the detection kit of the present invention and its use method
- the kit of the present invention detects the expression level of C1QBP in human oral mucosa tissue to determine the risk of oral squamous cell carcinoma. If the expression level of C1QBP is low (relative to healthy people), the risk of oral squamous cell carcinoma is low; if the expression level of C1QBP is high, the risk of oral mucosal squamous cell carcinoma is high, and the risk increases with the expression level of C1QBP. The higher the level, the worse the prognosis, that is, the longer the course of oral mucosal squamous cell carcinoma and the unsatisfactory outcome. It can be used for auxiliary diagnosis of clinical oral mucosal squamous cell carcinoma, providing effective basis for patients to take relevant treatment measures or decision-making, and has good clinical application prospects.
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Immunology (AREA)
- Molecular Biology (AREA)
- Chemical & Material Sciences (AREA)
- Biomedical Technology (AREA)
- Urology & Nephrology (AREA)
- Hematology (AREA)
- Cell Biology (AREA)
- Analytical Chemistry (AREA)
- Microbiology (AREA)
- General Health & Medical Sciences (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Biotechnology (AREA)
- Biochemistry (AREA)
- Pathology (AREA)
- General Physics & Mathematics (AREA)
- Hospice & Palliative Care (AREA)
- Oncology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
本发明涉及体外诊断试剂领域，具体涉及C1QBP蛋白在制备作为口腔癌筛查或预后标志物中的用途，以及检测C1QBP蛋白表达水平的试剂在制备口腔癌筛查或预后的试剂盒中的用途。本发明首次发现口腔鳞状细胞癌患者口腔黏膜组织中C1QBP蛋白表达水平显著高于健康患者。本发明将检测C1QBP蛋白表达水平的试剂用于制备口腔鳞状细胞癌筛查或预后的试剂盒，能够实现口腔鳞状细胞癌的有效筛查或预后。
Description
本发明涉及体外诊断试剂领域，具体涉及检测C1QBP蛋白表达水平的试剂在制备口腔癌筛查或预后试剂盒中的用途。
口腔鳞状细胞癌(OSCC)是临床最常见的口腔恶性肿瘤，尽管影像学、外科学、放疗和全身治疗的技术快速发展，但OSCC患者的总体生存率并没有明显的提高。治疗效果欠佳的主要原因之一包括淋巴结转移，而造成OSCC恶性转移生物学行为的发生机制尚不明确。目前研究表明，肿瘤细胞中存在复杂的代谢改变，肿瘤发生发展过程中存在的代谢异质性即肿瘤代谢重编程，肿瘤代谢重编程是恶性肿瘤的重要标志之一，与肿瘤生长、侵袭、转移及免疫逃逸等恶性行为密切相关，靶向代谢的肿瘤治疗具有的巨大临床应用潜力。
C1QBP又被称为gClqR、p32、p33和HABP1，C1QBP在可分布在细胞内、细胞表面，也可作为一种分泌蛋白被分泌到细胞外，在细胞内，C1QBP主要位于线粒体内，线粒体中C1QBP的重要功能是维持氧化磷酸化，为细胞提供能量并维持代谢平衡。在大量需要能量代谢的细胞(如心脏、骨骼肌、睾丸、卵巢、小肠和结肠中)的线粒体中，C1QBP均高表达。目前研究显示C1QBP在多种恶性肿瘤，例如乳腺癌、肺癌等中的表达增多，同时也有研究显示C1QBP在宫颈癌中低表达，说明C1QBP对肿瘤生物学行为的调节有组织特异性，但有关C1QBP在OSCC中表达和功能的研究尚未见报道。
发明内容
本发明的目的在于提供一种新的蛋白类的口腔癌标志物，以及该标志物的检测试剂在制备口腔癌筛查或预后的试剂盒中的用途。
本发明的技术方案包括：
C1QBP蛋白在制备作为口腔癌筛查或预后标志物中的用途。
检测C1QBP蛋白表达水平的试剂在制备口腔癌筛查或预后的试剂盒中的用途。
进一步地，所述检测C1QBP蛋白表达水平的试剂为酶联免疫吸附试验用试剂。
进一步地，所述检测C1QBP蛋白表达水平的试剂为免疫组织化学染色试验用试剂。
进一步地，所述口腔癌为口腔鳞状细胞癌。
更进一步地，所述检测C1QBP蛋白表达水平的试剂是检测人口腔黏膜组织中C1QBP蛋白表达水平的试剂。
本发明还提供了一种口腔癌筛查或预后的试剂盒，它包括用于检测C1QBP蛋白表达水平的试剂。
进一步地，所述检测C1QBP蛋白表达水平的试剂为酶联免疫吸附试验用试剂。
进一步地，所述检测C1QBP蛋白表达水平的试剂为免疫组织化学染色方法用试剂。
进一步地，所述口腔癌为口腔鳞状细胞癌。
更进一步地，所述检测C1QBP蛋白表达水平的试剂是检测人口腔黏膜组织中C1QBP蛋白表达水平的试剂。
本发明的关键在于，确定了人体口腔黏膜组织中C1QBP的表达水平与患口腔鳞状细胞癌的风险显著相关，因此可以通过检测人体口腔黏膜组织中C1QBP的表达水平来判断口腔鳞状细胞癌的风险，至于具体检测人体血液中C1QBP的表达水平手段，可以采用现有技术公开的各种手段，本发明实施例具体采用免疫组织化学染色方法进行检测，但不仅仅限于该手段，任何能够检测C1QBP的表达水平的方法均可用于口腔鳞状细胞癌筛查或预后。
本发明提供了一种新的口腔鳞状细胞癌筛查标记物和一种新的口腔鳞状细胞癌筛查试剂盒，能够实现口腔鳞状细胞癌的有效筛查和预测病程和结局，具备良好的应用前景。
显然，根据本发明的上述内容，按照本领域的普通技术知识和惯用手段，在不脱离本发明上述基本技术思想前提下，还可以做出其它多种形式的修改、替换或变更。
以下通过具体实施方式对本发明的上述内容再作进一步的详细说明。但不应将此理解为本发明上述主题的范围仅限于以下的实例。凡基于本发明上述内容所实现的技术均属于本发明的范围。
图1免疫组化染色强度评分标准
图2 C1QBP在口腔黏膜癌变各阶段的蛋白表达规律(A.C1QBP在正常口腔黏膜、口腔潜在恶性疾患(OPMD)和口腔鳞状细胞癌(OSCC)中的表达情况；B.C1QBP的表达随口腔黏膜癌变的进展变化情况)
图3 C1QBP的表达与OSCC患者预后的关系
实施例1口腔黏膜组织中C1QBP表达水平与口腔鳞状细胞癌的关系
一、方法
1、样本采集
首先收集正常口腔黏膜组织、口腔潜在恶性疾患(OPMD)患者病损组织、OSCC患者病损组织，其中正常对照组10例、OPMD组13例、OSCC组45例。将获得的组织按照下列方法进行固定、脱水、包埋，制作石蜡切片。
(1)收取的组织，立即在PBS中清洗两遍。
(2)轻轻用纸沾干组织，置于4％的多聚甲醛溶液中。组织与4％多聚甲醛液的体积比为1：30。
(3)根据分组，提前1天将固定的组织转入做好标记的包埋筐中，置于细流自来水下冲过夜。
(4)将包埋筐置于脱水机中脱水24h。
(5)取出包埋筐组织，经I蜡、II蜡、III蜡浸泡后，用包埋蜡包埋为蜡块。
(6)设置切片机为5μm厚度，切取组织石蜡切片，展平，载玻片捞片，标记好组别。
(7)置于55℃烤箱4h，烤干水分后保存于4℃冰箱中备用。
2、样本处理
将获得的组织切片按照下列方法进行免疫组织化学染色。
(1)石蜡切片置于70℃烤箱，1h。
(2)二甲苯及梯度酒精脱蜡：分别置于二甲苯I脱蜡20min，二甲苯II脱蜡10min。按照100％、95％、90％、80％和70％依次梯度酒精水化，每次5min。
(3)蒸馏水漂洗，5min，2次。
(4)抗原修复：准备高压锅加水，放置抗原修复盒，内装抗原修复液。本实验用枸橼酸钠缓冲液(10mM,pH6.0)。锅盖不严密闭合，火锅模式下加热至沸腾。将石蜡切片放入抗原修复盒中，盖紧盒盖，加热至上汽后计时3min。缓慢放气至无气体喷出，取下气阀，取出盒子。将盒子开盖，置于冷水下冷却20min。TBS漂洗，5min，2次。
(5)3％过氧化氢处理：准备30％的H2O2 20ml+蒸馏水180ml，石蜡切片置于H2O2中浸泡，室温避光放置20min。TBS清洗5min，3次。
(6)滴加Dako二抗显示系统A液，室温封闭30min，TBS清洗5min，3次。
(7)按稀释比滴加第一抗体，室温条件下孵育30min，然后置于4℃冰箱孵育过夜。
(8)过夜后取出，TBS洗5min，3次。
(9)滴加Dako二抗显示系统B液，37℃，孵育60min，TBS洗5min，3次。
(10)每张切片滴加DAB显色液(1:50稀释)并置于显微镜下观察，片子刚显示棕黄色时，放入清水中终止。
(11)苏木素复染1min，蒸馏水冲洗，2％盐酸酒精分色1s，蒸馏水清洗15min，氨水返蓝。
(12)洁净水细流冲洗20min。
(13)梯度酒精脱水干燥(依次过75％酒精、80％酒精、90％酒精和100％酒精，每次5min)，二甲苯透明(依次过二甲苯I和二甲苯II，每次5min)。
(14)中性树胶封片，自然晾干。
(15)扫描切片，光盘刻录图像数据。
3.结果统计
使用高倍镜观察每张切片上5个不重复的视野，C1QBP以细胞浆中出现黄色或棕黄色颗粒作为阳性，分别评估整张切片的平均染色强度以及在每个视野中阳性细胞占肿瘤细胞的百分比进行免疫组织化学染色评分(IRS)。所有切片均经过两名有经验的研究员进行盲测，评估并计算出每张切片的平均分。评分标准如下(表1和图1)
表1.IHC染色评分标准
4、结果分析
通过统计分析，正常口腔黏膜组织C1QBP的表达水平相对较低(平均分为0.900)、口腔潜在恶性疾患(OPMD)患者病损组织C1QBP的表达水平增强(平均分为3.538)，OSCC患者组织C1QBP的表达水相对最强(平均分为6.067)，OPMD组与正常口腔黏膜组相比有统计学意义(P<0.05)，OSCC组与正常口腔黏膜组相比有统计学意义(P<0.001)，OSCC组与OPMD组相比有统计学意义(P<0.01)；C1QBP的表达水平与OSCC患者的预后呈负相关关系，差异均具有统计学意义(P<0.05)。
由上述结果可知，C1QBP的表达水平随口腔黏膜癌变的进展而升高(图2)，且与OSCC患者的预后呈负相关关系(图3)，通过检测口腔黏膜组织中C1QBP表达水平，能够达到口腔鳞状细胞癌筛查或预测口腔鳞状细胞癌疾病的未来病程和结局的目的。
实施例2本发明的检测试剂盒的组成及其使用方法
一、试剂盒组成
二、试剂盒使用方法
同实施例1“石蜡切片制作”和“免疫组织化学染色”。
本发明的试剂盒检测人体口腔黏膜组织中C1QBP的表达水平来判断口腔鳞状细胞癌的风险。若C1QBP的表达水平低(相对于健康人而言)，则患口腔鳞状细胞癌的风险低；若C1QBP的表达水平高，则患口腔黏膜鳞状细胞癌风险高，且随C1QBP的表达水平升高，预后越差，即口腔黏膜鳞状细胞癌的病程越长，结局不理想。可用于临床口腔黏膜鳞状细胞癌的辅助诊断，为患者采取相关的治疗措施或者决策提供有效的依据，临床应用前景良好。
Claims (10)
- C1QBP蛋白在制备作为口腔癌筛查或预后标志物中的用途。
- 检测C1QBP蛋白表达水平的试剂在制备口腔癌筛查或预后的试剂盒中的用途。
- 如权利要求2所述的用途，其特征在于，所述检测C1QBP蛋白表达水平的试剂为酶联免疫吸附试验用试剂。
- 如权利要求2所述的用途，其特征在于，所述检测C1QBP蛋白表达水平的试剂为免疫组织化学染色试验用试剂。
- 如权利要求2所述的用途，其特征在于，所述口腔癌为口腔鳞状细胞癌。
- 如权利要求2～5任一所述的用途，其特征在于，所述检测C1QBP蛋白表达水平的试剂是检测人口腔黏膜组织中C1QBP蛋白表达水平的试剂。
- 一种口腔癌筛查或预后的试剂盒，其特征在于，它包括用于检测C1QBP蛋白表达水平的试剂。
- 如权利要求7所述的试剂盒，其特征在于，所述检测C1QBP蛋白表达水平的试剂为酶联免疫吸附试验用试剂或免疫组织化学染色方法用试剂。
- 如权利要求7所述的试剂盒，其特征在于，所述口腔癌为口腔鳞状细胞癌。
- 如权利要求7～9任一所述的试剂盒，其特征在于，所述检测C1QBP蛋白表达水平的试剂是检测人口腔黏膜组织中C1QBP蛋白表达水平的试剂。
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210499870.2 | 2022-05-09 | ||
| CN202210499870.2A CN117074683A (zh) | 2022-05-09 | 2022-05-09 | 检测c1qbp蛋白表达水平的试剂在制备口腔癌筛查或预后试剂盒中的用途 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| WO2023217035A1 true WO2023217035A1 (zh) | 2023-11-16 |
Family
ID=88704840
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| PCT/CN2023/092512 WO2023217035A1 (zh) | 2022-05-09 | 2023-05-06 | 检测c1qbp蛋白表达水平的试剂在制备口腔癌筛查或预后试剂盒中的用途 |
Country Status (2)
| Country | Link |
|---|---|
| CN (1) | CN117074683A (zh) |
| WO (1) | WO2023217035A1 (zh) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104093415A (zh) * | 2011-10-24 | 2014-10-08 | 哈洛齐梅公司 | 抗乙酰透明质酸剂疗法的同伴诊断剂及其使用方法 |
| CN104267191A (zh) * | 2014-09-09 | 2015-01-07 | 北京大学口腔医学院 | 口腔口咽鳞状细胞癌的生物标志物及其应用 |
| CN107422123A (zh) * | 2017-07-26 | 2017-12-01 | 复旦大学附属中山医院 | 一种用于诊断口腔鳞状细胞癌的试剂盒 |
| CN112512570A (zh) * | 2018-02-27 | 2021-03-16 | 迪亚库雷特公司 | 疗法中的pla2-gib调节 |
-
2022
- 2022-05-09 CN CN202210499870.2A patent/CN117074683A/zh active Pending
-
2023
- 2023-05-06 WO PCT/CN2023/092512 patent/WO2023217035A1/zh unknown
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104093415A (zh) * | 2011-10-24 | 2014-10-08 | 哈洛齐梅公司 | 抗乙酰透明质酸剂疗法的同伴诊断剂及其使用方法 |
| CN104267191A (zh) * | 2014-09-09 | 2015-01-07 | 北京大学口腔医学院 | 口腔口咽鳞状细胞癌的生物标志物及其应用 |
| CN107422123A (zh) * | 2017-07-26 | 2017-12-01 | 复旦大学附属中山医院 | 一种用于诊断口腔鳞状细胞癌的试剂盒 |
| CN112512570A (zh) * | 2018-02-27 | 2021-03-16 | 迪亚库雷特公司 | 疗法中的pla2-gib调节 |
Non-Patent Citations (3)
| Title |
|---|
| FOGAL VALENTINA, ZHANG LIANGLIN, KRAJEWSKI STAN, RUOSLAHTI ERKKI: "Mitochondrial/Cell-Surface Protein p32/gC1qR as a Molecular Target in Tumor Cells and Tumor Stroma", CANCER RESEARCH, AMERICAN ASSOCIATION FOR CANCER RESEARCH, US, vol. 68, no. 17, 1 September 2008 (2008-09-01), US, pages 7210 - 7218, XP093108370, ISSN: 0008-5472, DOI: 10.1158/0008-5472.CAN-07-6752 * |
| WANG DAWEI, ZHANG JINTING, LI QINGXING: "Expression and Clinical Significance of P33ING1b Protein in Oral Squamous Cell Carcinoma", JOURNAL OF PRACTICAL STOMATOLOGY, vol. 22, no. 06, 30 November 2006 (2006-11-30), pages 811 - 813, XP009550377, ISSN: 1001-3733 * |
| WANG ZHENG, SUN ANQI, YAN AIHUI, YAO JIAN, HUANG HAIBO, GAO ZIMING, HAN TAO, GU JIA, LI NI, WU HUIZHE, LI KAI: "Circular RNA MTCL1 promotes advanced laryngeal squamous cell carcinoma progression by inhibiting C1QBP ubiquitin degradation and mediating beta-catenin activation", MOLECULAR CANCER, BIOMED CENTRAL, LONDON, GB, vol. 21, no. 1, GB , XP093108375, ISSN: 1476-4598, DOI: 10.1186/s12943-022-01570-4 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN117074683A (zh) | 2023-11-17 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Decenzo et al. | Antigenic deletion and prognosis of patients with stage A transitional cell bladder carcinoma | |
| Sofi et al. | Estrogen receptor and progesterone receptor status in breast cancer in relation to age, histological grade, size of lesion and lymph node involvement | |
| CN110187110B (zh) | 一种贲门癌预后预测标志物及其应用 | |
| CN108369234B (zh) | 基于标志物分子将个体鉴定为待通过化疗治疗的方法和相关用途 | |
| CN110836974B (zh) | Flt3lg蛋白在制备肺腺癌术后预后评估试剂或者试剂盒中的应用 | |
| CN110283909A (zh) | Zbtb20蛋白或其特异性抗体在贲门癌检测试剂盒中的应用 | |
| CN113834941B (zh) | 基于b细胞表达的结肠癌预后诊断用标记物及其用途 | |
| WO2023217035A1 (zh) | 检测c1qbp蛋白表达水平的试剂在制备口腔癌筛查或预后试剂盒中的用途 | |
| CN113970638B (zh) | 确定胃癌极早期发生风险及评估胃癌前病变进展风险的分子标志及其在诊断试剂盒中的应用 | |
| CN111856014A (zh) | 诊断和治疗膀胱癌的分子标志物mllt11及其用途 | |
| MOHSENI et al. | Prognostic values of proliferative markers ki-67 and repp86 in breast cancer | |
| CN113238052B (zh) | MG7-Ag、hTERT及TFF2表达分析在肠上皮化生风险分层及胃癌预警中的应用 | |
| CN111551720A (zh) | 一种检测ogn蛋白表达的方法及其在胃癌辅助诊断中的应用 | |
| CN107422123B (zh) | 一种用于诊断口腔鳞状细胞癌的试剂盒 | |
| CN111665358B (zh) | Nalcn蛋白在食管鳞癌预后预测中的应用 | |
| Liu et al. | Localization of indoleamine 2, 3-dioxygenase in human esophageal squamous cell carcinomas | |
| CN105785004B (zh) | 细胞分裂周期相关蛋白2在胰腺癌诊断或预后中的用途 | |
| CN112229997B (zh) | 一种卵巢癌的预后诊断标志物Claudin23及其应用 | |
| Shui et al. | Relationship between cyclooxygenase-2 (COX-2) content and prognosis in nasopharyngeal carcinoma before and after radiochemotherapy | |
| CN107817350A (zh) | 一种肺癌筛查试剂盒 | |
| CN106680510B (zh) | Myoferlin及其特异性抗体在制备检测鼻咽癌试剂盒中的应用 | |
| Momin et al. | Expression Of P63 In Oral Dysplastic And Malignant Squamous Cell Carcinoma | |
| CN102297971A (zh) | 一种检测癌或恶性肿瘤组织中jwa表达的试剂盒 | |
| CN110095606B (zh) | 一种肺癌的筛查试剂盒 | |
| CN117368479B (zh) | 一种用于肺腺癌诊断的生物标志物及检测试剂盒 |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| 121 | Ep: the epo has been informed by wipo that ep was designated in this application |
Ref document number: 23802807 Country of ref document: EP Kind code of ref document: A1 |