Method for diagnosis of bile duct cancer using methionyl-trna synthetase in bile duct cell

A technology of methionyl and bile duct cells, applied in the field of diagnosis of cholangiocarcinoma, can solve the problems of no markers and pathological diagnosis methods

Pending Publication Date: 2020-04-10
温口特
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The real problem is that even though the cancer is actually progressing, in most cases there are only findings of atypical cells or tissues on examination at the tissue or cellular level
[0011] Despite these studies and efforts in the diagnosis of biliary tract cancer, there are still no effective markers and pathological diagnostic methods that can accurately distinguish and determine malignant and Benign state (biliary tract disease other than cholangiocarcinoma)

Method used

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  • Method for diagnosis of bile duct cancer using methionyl-trna synthetase in bile duct cell
  • Method for diagnosis of bile duct cancer using methionyl-trna synthetase in bile duct cell
  • Method for diagnosis of bile duct cancer using methionyl-trna synthetase in bile duct cell

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Experimental program
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Effect test

Embodiment 1

[0136] Example 1: The present invention is used for the construction of the antibody of cholangiocarcinoma inspection (obtaining the antibody with high specificity to MRS)

[0137] It is known that in vivo, methionyl-tRNA synthetase (MRS) exists in a state bound to aminoacyl-tRNA synthetase complex-interacting multifunctional protein 3 (AIMP3), and this binding is disrupted by UV irradiation or the like. Thus, for substantially accurate detection of MRS, only the detection of MRS specifically is required, even in the case of MRS binding to AIMP3. However, the protein structures between current AIMP types and ARS types have many similarities, so commercially available antibodies have the problem of showing cross-activity with different AIMP and ARS types. For the diagnostic accuracy of the cholangiocarcinoma examination method of the present invention, the present inventors prepared a highly sensitive MRS antibody that has no cross activity with other proteins as follows.

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Embodiment 2

[0179] Example 2: Establishment of a specific MRS expression detection method (staining method) for cholangiocarcinoma cells in cell diagnosis and confirmation of its effect

[0180] 1) Obtain specimens according to commonly used brush cytology (Osnes M, Serck-Hanssen A, Myren J. Scand J Gastroenterol. 1975; 10(8):829-31). Specifically, bile duct brushing was performed using a GRBH-230-3-3.5 brush (Wilson-Cook Medical, Inc., Winston-Salem, NC). Allow the brush to move back and forth over the lesion five to eight times. Afterwards, the brushes were washed with Roswel Park Memorial Institute (RPMI) 1640 medium (GibcoBRL, Rockville, MA, USA) and immediately moved to the cytology laboratory for liquid-based cytology (Thinprep). Shake the brush in ThinPrep fixative solution (PreservvCyt solution) to release cholangiocytes. The sample thus obtained (cholangiocytes) was spread on a ThinPrep glass slide by a conventional method using ThinPrep (Hologic Inc) to prepare a cell sample. ...

Embodiment 3

[0203] Example 3: In biopsy, the establishment of a specific MRS expression detection method for cholangiocarcinoma cells and the confirmation of its effect

[0204] Immunohistochemistry (IHC) for determining the extent of MRS expression in cholangiocarcinoma tissue and normal bile duct tissue (including benign bile duct stricture cells but not cancer) was developed as follows. Specifically, typically 55 unknown bile duct biopsy tissue samples were paraffin-embedded and sectioned. Afterwards, the final samples are obtained by processing in the following order:

[0205] ① Soak sliced ​​tissue in xylene for 24 hours → treat with 100% alcohol twice for 2 minutes → treat with 95% alcohol once for 2 minutes → treat with 90% alcohol twice for 2 minutes → treat with 70% alcohol once for 2 minutes → wash with DW two or three times;

[0206] ② Antigen recovery: dilute commercially available citrate buffer (DW 9: citrate buffer 1), preheat for 2 minutes, put slides in it, and heat for...

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Abstract

The present invention relates to a method for diagnosis of bile duct cancer, using methionyl-tRNA synthetase (MRS) in bile duct cells and, more particularly, to a composition for diagnosing bile ductcancer, comprising an agent with which an expression level of methionyl-tRNA synthetase protein is measured, a diagnostic kit, and a method for qualitative or quantitative analysis of MRS to provide information necessary for the diagnosis of bile duct cancer. According to the present invention, MRS retains a very high value as a diagnostic marker for bile duct cancer in terms of rapidity and accuracy because MRS allows determination to be made to see whether bile duct cancer is present or absent for bile duct cells, which have been classified as atypical cells by conventional pathological examination methods. Hence, the present invention provides a method for discriminating between cancer cells and normal cells in atypical cells and can make a definite diagnosis of cancer with almost 100 %in all of sensitivity, specificity, and accuracy in contrast to many conventional cancer markers with which an actual result of diagnosis is substantially difficult to obtain at a cell level (that is, cytodiagnosis).

Description

technical field [0001] The present invention relates to a method for diagnosing cholangiocarcinoma by using methionyl-tRNA synthetase (MRS) in cholangiocytes. More specifically, the present invention relates to a composition for diagnosing cholangiocarcinoma, said composition comprising reagents for determining the expression level of methylsulfonyl-tRNA synthetase protein; to a diagnostic kit; and to a A method for qualitative or quantitative analysis of MRS to provide the information required for the diagnosis of cholangiocarcinoma. Background technique [0002] This application claims priority from Korean Patent Application No. 10-2017-0059318 filed on May 12, 2017, the entire contents of which are incorporated herein by reference. [0003] The incidence rate of cholangiocarcinoma is 6.3 cases per 100,000 population, ranking 9th among all cancers, but its prognosis is poor, ranking 6th among the causes of cancer death in Korea. Until a definitive diagnosis of cholangioc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/574G01N33/68
CPCG01N33/57407G01N33/57492G01N33/6893C12Y601/0101C07K16/40C07K2317/56C12N9/93G01N1/30G01N33/5091G01N33/573
Inventor 金圣勋权南勋李东起林氾镇张星日
Owner 温口特
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