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Applications of Galectin-10 and specific antibody thereof in preparation of kit for detecting nasopharyngeal carcinoma

A kit and technology for nasopharyngeal carcinoma, applied in the field of diagnostic markers, can solve problems such as biological functions that have not been reported yet

Active Publication Date: 2017-05-10
XIANGYA HOSPITAL CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It has been reported that Galectin-10 is involved in the regulation of cell growth, angiogenesis and inflammation progression, but its biological function in nasopharyngeal carcinoma has not been reported yet

Method used

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  • Applications of Galectin-10 and specific antibody thereof in preparation of kit for detecting nasopharyngeal carcinoma
  • Applications of Galectin-10 and specific antibody thereof in preparation of kit for detecting nasopharyngeal carcinoma
  • Applications of Galectin-10 and specific antibody thereof in preparation of kit for detecting nasopharyngeal carcinoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] This example is used to illustrate the clinical significance of immunohistochemical analysis of the expression of Galectin-10 in nasopharyngeal carcinoma tissues:

[0023] Galectin-10 specific antibody (purchased by sigma aldrich, C0998) combined with Maixin S-P immunohistochemistry kit (Fujian Maixin Company) constitutes the diagnostic kit of this embodiment. Biopsy nasopharyngeal tissues were fixed with 10% paraformaldehyde, dehydrated and transparent with conventional graded alcohol, embedded in paraffin, cut into 4 μm thick slices, and baked. Perform immunohistochemical staining according to the instructions of the immunohistochemical hypersensitivity kit:

[0024] 1) Dewax the paraffin tissue sections twice with 100% xylene, half an hour each time, until completely dewaxed;

[0025] 2) After hydration with freshly prepared 100%, 95%, 90%, 75% gradient alcohol, rinse with PBS solution (PH6.0) 3 times, 3 min each time;

[0026] 3) Antigen retrieval: place the treat...

Embodiment 2

[0043] This example is used to illustrate the expression of Galectin-10 protein detected by Western blotting

[0044]Protein preparation: Nasopharyngeal carcinoma cell lines CNE1, CNE2 and HK1 were routinely cultured with RPMI 1640 complete medium containing 100 U / ml antibiotics (penicillin and streptomycin) and 10% calf serum (37°C, 5% CO2, saturated humidity conditions). NP69 cells were cultured with 1×KSFM special medium containing 10% calf serum and 100U / ml antibiotics. When all cells grow to about 80% confluence, wash the cell surface three times with PBS, collect about 1×107 cells by centrifugation, add 400 μL of modified RIPA lysate (50mmol / L Tris HCl [pH 8.0], 150mmol / L NaCl, 0.5% Sodium deoxycholate, 1mmol / LEDTA 1%, 1% NP-40, 0.1μg / ml PMSF, 2μg / ml leustatin), mixed thoroughly, lysed on ice for 30min, centrifuged at 12000rpm at 4°C for 30min, and sucked the supernatant as the total protein. Determination of protein concentration by Bradford method: add 2 mg / ml bovin...

Embodiment 3

[0047] Example 3 ELISA detection of the concentration of Galectin-10 in nasopharyngeal carcinoma serum

[0048] In order to verify the difference of Galectin-10 in the serum of human nasopharyngeal carcinoma and adjacent tumors, the ELISA method was used in this example to detect the expression level of Galectin-10 in the serum of 28 patients with nasopharyngeal carcinoma and 17 normal subjects.

[0049] The operation process is as follows:

[0050] 1) Collect about 500 μL of nasopharyngeal carcinoma or normal human serum and store it at -80°C for later use.

[0051] 2) Purchase human crystal protein (Charcot Leyden Crystal Protein, CLC, also known as Galectin-10) enzyme-linked immunosorbent assay kit (product number: E-EL-H1517c) from Elabscience Biotechnology Company, and detect Galectin-10 in serum according to the operation instructions concentration.

[0052] 3) Adding samples: set blank wells, standard wells, and sample wells to be tested respectively. Add 100 μL of s...

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Abstract

The invention relates to a diagnostic marker, particularly discloses a nasopharyngeal carcinoma related marker Galectin-10 (genebank accession number: ID1178), and provides applications of the marker and a specific antibody thereof in preparation of a kit for detecting nasopharyngeal carcinoma, wherein the kit can be an immunohistochemical kit or an enzyme-linked immunosorbent assay kit. The invention provides a new scientific basis for effectively judging human nasopharyngeal disease progression, and can achieve early discovery and early treatment of the nasopharyngeal carcinoma.

Description

technical field [0001] The present invention relates to a diagnostic marker, in particular to the application of Galectin-10 in detecting nasopharyngeal carcinoma. Background technique [0002] Nasopharyngeal carcinoma is a malignant tumor of epithelial origin that frequently occurs in southern provinces of my country and Southeast Asia with significant ethnic and geographical distribution characteristics. The pathogenesis of nasopharyngeal carcinoma is related to genetic factors, Epstein-Barr virus infection and other environmental factors. [0003] With the rapid development of medical technology, more and more advanced equipment has been used in the examination of nasopharyngeal carcinoma. At present, the detection methods of nasopharyngeal carcinoma mainly include clinical examination, nasopharyngoscopy, imaging examination, pathological examination and serological examination. [0004] Imaging examinations have limited sensitivity and are expensive; pathological exami...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/574
CPCG01N33/57407G01N33/6893G01N2333/46G01N2800/14
Inventor 汤参娥彭芳朱江赵师刘芬李巧羽朱劲飞
Owner XIANGYA HOSPITAL CENT SOUTH UNIV
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