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Esophageal cancer marker and use thereof

a technology for esophageal cancer and tumor markers, applied in the field of esophageal cancer, can solve the problem of not having a definitive tumor marker for esophageal cancer

Inactive Publication Date: 2018-12-20
NAT UNIV CORP KOCHI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0052]According to the present invention, an effective marker for esophageal cancer is provided, and further it was found that the marker can be utilized in the treatment or prevention of esophageal cancer. Therefore, it is possible to carry out diagnosis, treatment, or prevention of esophageal cancer in an early stage, which was previously impossible or difficult.

Problems solved by technology

However, as of present, the tumor markers are used to understand the dynamics of progressing malignant tumor is understood, but a definitive tumor marker for esophageal cancer is not available.

Method used

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  • Esophageal cancer marker and use thereof
  • Esophageal cancer marker and use thereof
  • Esophageal cancer marker and use thereof

Examples

Experimental program
Comparison scheme
Effect test

example 1

n of Glypican-1 in Various Cells by Western Blot

[0326]In the present example, the expression of Glypican-1 in various cells by Western blot was investigated.

(Western Blot Analysis)

[0327]Normal esophageal epithelial cells HEEpic and Het1A and esophageal squamous cell carcinoma cell strains TE1, TE5, TE6, TE8, TE9, TE10, TE11, TE14, and TE15 were washed with ice-cooled PBS (−), and then peeled off with a cell scraper. The cells were then recovered by centrifugation. The cells were lysed by a Lysis buffer (20 mM Tris-HCl, pH 7.5, 150 mM NaCl, 1% Triton X-100, 1× protease inhibitor cocktail (NACALAI TESQUE), 1× phosphatase inhibitor cocktail (NACALAI TESQUE)), The supernatant was recovered as a protein extract liquid by centrifugation (13,200 rpm, 4° C., 15 min). The protein concentration was quantified with a protein quantifying kit (DC Protein Assay kit (Bio-Rad Laboratories, Inc.)) using bovine serum albumin (BSA) as a standard.

[0328]A suger chain of Glypican-1 was enzymatically clea...

example 2

Expression Level of Glypican-1 (Accession No. P35052)

[0332]In the present example, the relative expression level of Glypican-1 (Accession No. P35052) was investigated in normal cells and various esophageal cancer cell strains.

(Techniques)

[0333]The expression level of Glypican-1 in esophageal squamous cell carcinoma cell strains TE1, TE6, TE8, TE9, TE10, and TE14 was evaluated relative to normal esophageal epithelial cells HEEpic and Het1A.

[0334]For 8 types of cell strains cultured in 150 mm Petri dishes, a cell surface membrane protein comprising Glypican-1 was biotinylated with sulfo-NHS-SS-biotin. The extracted protein was purified by Neurto-avidin beads. At that time, in order to correct for the error among the samples, sulfo-NHS-SS-biotin-labeled bovine serum albumin was added to each in an equal amount as an internal standard, and was used for correction of quantification results from a mass spectrometer. The purified proteins were digested by trypsin and labeled with an iTRAQ ...

example 3

ysis to Show that Glypican-1 is Expressed on Cell Surface of Esophageal Cancer Cells

[0336]Then, in the present example, it was confirmed by FACS that Glypican-1 was expressed on cell surface of esophageal cancer cells.

(Facs Analysis)

[0337]Cells were washed with PBS (Nacalai Tesque) twice and peeled off from a dish by 0.02% EDTA solution (Nacalai Tesque). The cells were washed with a FACS staining buffer (PBS supplemented with 1% FBS and 0.1% sodium azide) twice, stained with a 5-time-diluted goat anti-human Glypican-1 antibody (R&D Systems, Minneapolis, Minn.), and subsequently stained with a 50-time-diluted PE-labeled anti-goat IgG antibody. The stained cells were measured by FACS Canto II (Becton Dickinson, Mountain View, Calif., USA) and the data was analyzed using FlowJo software (TreeStar, Stanford, Calif., USA).

(Result)

[0338]The result is shown in FIG. 3. As shown, while the expression of Glypican-1 in the normal cells was considered the background, it was shown that in all th...

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Abstract

The present invention relates to an esophageal cancer marker and application thereof. The present invention relates to: a marker that includes Glypican-1 or an expression product thereof, or a fragment or derivative thereof, and serves to identify esophageal cancer; a detection agent that includes a substance that binds to Glypican-1 or an expression product thereof; and a composition that includes a Glypican-1 inhibitor and serves to prevent or treat esophageal cancer. Herein, Glypican-1 can be SEQ ID NO: 1 (nucleic acid sequence) or SEQ ID NO: 2 (amino acid sequence), or an equivalent thereof.

Description

STATEMENT REGARDING SEQUENCE LISTING[0001]The Sequence Listing associated with this application is provided in text format in lieu of a paper copy, and is hereby incorporated by reference into the specification. The name of the text file containing the Sequence Listing is 691188_403D1_SEQUENCE_LISTING.txt. The text file is 54.2 KB, was created on Aug. 13, 2018, and is being submitted electronically via EFS-Web.TECHNICAL FIELD[0002]The present invention is related to markers for cancer, in particularly esophageal cancer, and relevant technologies, methods, agents, and the like.BACKGROUND ART[0003]Esophageal cancer is cancer occurring in the esophagus and subjective symptoms often do not appear in the early stage thereof, while esophageal cancer found with no symptoms is in the early stage and thus the probability of curing is high. On the other hand, as tumor markers for esophageal cancer, SCC (squamous cell carcinoma related antigen), CEA (carcinoembryonic antigen), CA19-9, p53, and...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C07K16/30C12N15/113C07K16/28G01N33/574C12Q1/6886A61K31/713A61K39/00
CPCC12Q1/6886C12Q2600/158G01N33/57407G01N2333/705C07K2317/565C07K2317/76C07K16/30C07K2317/33C12N15/1138C12N2310/14C07K2317/92C07K2317/73C07K2317/622C07K2317/23A61K2039/505C07K16/28A61K31/713G01N2400/40A61P1/00A61P1/04A61P35/00A61P35/04A61P43/00
Inventor NAKA, TETSUJISERADA, SATOSHIFUJIMOTO, MINORUTOYOURA, MASAYOSHISHOYA, YUJI
Owner NAT UNIV CORP KOCHI UNIV
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