a 2023

Verification of diagnosis using CDX2 gene expression in esophageal fresh frozen tissue

MAJER, Adam, Natálie MLČŮCHOVÁ, Zdeněk PAVLOVSKÝ, Lumír KUNOVSKÝ, Radek KROUPA et. al.

Basic information

Original name

Verification of diagnosis using CDX2 gene expression in esophageal fresh frozen tissue

Name (in English)

Verification of diagnosis using CDX2 gene expression in esophageal fresh frozen tissue

Authors

MAJER, Adam (203 Czech Republic), Natálie MLČŮCHOVÁ (203 Czech Republic), Zdeněk PAVLOVSKÝ (203 Czech Republic), Lumír KUNOVSKÝ (203 Czech Republic), Radek KROUPA (203 Czech Republic), Zdeněk KALA (203 Czech Republic) and Petra BOŘILOVÁ LINHARTOVÁ (203 Czech Republic, guarantor, belonging to the institution)

Edition

Setkání biochemiků a molekulárních biologů, 2023

Other information

Language

Czech

Type of outcome

Konferenční abstrakt

Field of Study

10608 Biochemistry and molecular biology

Country of publisher

Czech Republic

Confidentiality degree

není předmětem státního či obchodního tajemství

RIV identification code

RIV/00216224:14310/23:00133553

Organization unit

Faculty of Science

Keywords in English

gene expression; immunohistochemistry; CDX2; Barrett’s esophagus; adenocarcinoma
Změněno: 16/2/2024 13:19, Mgr. Terezie Slámová

Abstract

V originále

Introduction The fresh frozen tissue (FFT) is preferably used for some molecular analyses, such as metagenomics and metatranscriptomics. In general, an examination should be performed to determine whether the bioptic tissue is pathological. CDX2 is expressed by intestinal cells and is already used as a diagnostic marker for Barrett’s esophagus (BE) and esophageal adenocarcinoma (EAC). This study aims to evaluate CDX2 gene expression in esophageal FFT from patients with gastroesophageal reflux disease (GERD) and to compare findings with histopathological diagnosis of parallel samples. Methods In 23 patients with GERD, endoscopic examination was carried out and 92 esophageal and 3 duodenal biopsies were taken. Two parallel bioptic tissues were obtained from two different sites of esophageal mucosa – site with the main pathology and adjacent site with macroscopically normal mucosa. 46 paired formalin-fixed paraffin-embedded (FFPE) samples were examined by pathologist and CDX2-staining immunohistochemistry (IHC) was performed. RNA was extracted from 46 paired esophageal FFT and 3 duodenum samples (served as positive controls) using the AllPrep DNA/RNA Mini Kit. Reverse transcription was performed using High-Capacity cDNA Reverse Transcription Kit and CDX2 mRNA levels were assessed using qPCR and Taqman Gene Expression Assays. Results All BE and EAC FFPE specimens revealed immunoreactivity for CDX2, while all esophageal FFPE samples from adjacent site were IHC-negative for CDX2. The CDX2 was expressed in 91 % of esophageal FFT with the main pathology from BE patients, in 58 % of esophageal FFT with the main pathology from EAC patients, as well as in 4.3 % of esophageal FFT taken from the macroscopically normal esophageal mucosa. Conclusion This study shows that a percentage of esophageal biopsies taken for molecular analyses has different diagnosis based on the CDX2 gene expression than parallel biopsies examined histopathollogically. For further metagenome or metatranscriptome studies, there is a need to find out a suitable marker, such as CDX2 in the case of BE and EAC, and to examine whether the tissue really represents the pathological or physiological condition.

In English

Introduction The fresh frozen tissue (FFT) is preferably used for some molecular analyses, such as metagenomics and metatranscriptomics. In general, an examination should be performed to determine whether the bioptic tissue is pathological. CDX2 is expressed by intestinal cells and is already used as a diagnostic marker for Barrett’s esophagus (BE) and esophageal adenocarcinoma (EAC). This study aims to evaluate CDX2 gene expression in esophageal FFT from patients with gastroesophageal reflux disease (GERD) and to compare findings with histopathological diagnosis of parallel samples. Methods In 23 patients with GERD, endoscopic examination was carried out and 92 esophageal and 3 duodenal biopsies were taken. Two parallel bioptic tissues were obtained from two different sites of esophageal mucosa – site with the main pathology and adjacent site with macroscopically normal mucosa. 46 paired formalin-fixed paraffin-embedded (FFPE) samples were examined by pathologist and CDX2-staining immunohistochemistry (IHC) was performed. RNA was extracted from 46 paired esophageal FFT and 3 duodenum samples (served as positive controls) using the AllPrep DNA/RNA Mini Kit. Reverse transcription was performed using High-Capacity cDNA Reverse Transcription Kit and CDX2 mRNA levels were assessed using qPCR and Taqman Gene Expression Assays. Results All BE and EAC FFPE specimens revealed immunoreactivity for CDX2, while all esophageal FFPE samples from adjacent site were IHC-negative for CDX2. The CDX2 was expressed in 91 % of esophageal FFT with the main pathology from BE patients, in 58 % of esophageal FFT with the main pathology from EAC patients, as well as in 4.3 % of esophageal FFT taken from the macroscopically normal esophageal mucosa. Conclusion This study shows that a percentage of esophageal biopsies taken for molecular analyses has different diagnosis based on the CDX2 gene expression than parallel biopsies examined histopathollogically. For further metagenome or metatranscriptome studies, there is a need to find out a suitable marker, such as CDX2 in the case of BE and EAC, and to examine whether the tissue really represents the pathological or physiological condition.

Links

LM2023069, research and development project
Name: Výzkumná infrastruktura RECETOX
Investor: Ministry of Education, Youth and Sports of the CR, RECETOX research infrastructure
NU20-03-00126, research and development project
Name: Hostitelský mikrobiom ve vztahu k rozvoji Barrettova jícnu a adenokarcinomu jícnu
Investor: Ministry of Health of the CR, Host microbiome in relation to Barrett ́s esophagus and esophageal adenocarcinoma development, Subprogram 1 - standard