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.

Základní údaje

Originální název

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

Název anglicky

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

Autoři

MAJER, Adam (203 Česká republika), Natálie MLČŮCHOVÁ (203 Česká republika), Zdeněk PAVLOVSKÝ (203 Česká republika), Lumír KUNOVSKÝ (203 Česká republika), Radek KROUPA (203 Česká republika), Zdeněk KALA (203 Česká republika) a Petra BOŘILOVÁ LINHARTOVÁ (203 Česká republika, garant, domácí)

Vydání

Setkání biochemiků a molekulárních biologů, 2023

Další údaje

Jazyk

čeština

Typ výsledku

Konferenční abstrakt

Obor

10608 Biochemistry and molecular biology

Stát vydavatele

Česká republika

Utajení

není předmětem státního či obchodního tajemství

Kód RIV

RIV/00216224:14310/23:00133553

Organizační jednotka

Přírodovědecká fakulta

Klíčová slova anglicky

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

Anotace

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.

Anglicky

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.

Návaznosti

LM2023069, projekt VaV
Název: Výzkumná infrastruktura RECETOX
Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, Výzkumná infrastruktura RECETOX
NU20-03-00126, projekt VaV
Název: Hostitelský mikrobiom ve vztahu k rozvoji Barrettova jícnu a adenokarcinomu jícnu
Investor: Ministerstvo zdravotnictví ČR, Hostitelský mikrobiom ve vztahu k rozvoji Barrettova jícnu a adenokarcinomu jícnu, Podprogram 1 - standardní