Proteomic Screening of a Cell Line Model of Esophageal Carcinogenesis Identifies Cathepsin D and Aldo-Keto Reductase 1C2 and 1B10 Dysregulation in Barrett’s Esophagus and Esophageal Adenocarcinoma

Abstract

Esophageal adenocarcinoma (EA) incidence is increasing rapidly and is associated with a poor prognosis. Identifying biomarkers of disease development and progression would be invaluable tools to inform clinical practice. Two-dimensional polyacrylamide gel electrophoresis was used to screen 10 esophageal cell lines representing distinct stages in the development of esophageal cancer. Thirty-three proteins were identified by MALDI-TOF-MS which demonstrated differences in expression across the cell lines. Western blotting and qRT-PCR confirmed increased cathepsin D and aldo-keto reductases 1C2 and 1B10 expression in metaplastic and dysplastic cell lines. Expression of these proteins was further assessed in esophageal epithelium from patients with nonerosive (NERD) and erosive gastro-esophageal reflux disease, Barrett's esophagus (BE) and EA. When compared with normal epithelium of NERD patients, (i) cathepsin D mRNA levels demonstrated a stepwise increase in expression (p<0.05) in erosive, metaplastic and EA tissue; (ii) AKR1B10 expression increased (p<0.05) 3- and 9-fold in erosive and Barrett's epithelium, respectively; and (iii) AKR1C2 levels increased (p<0.05) in erosive and Barrett's epithelium, but were reduced (p<0.05) in EA. These proteins may contribute to disease development via effects on apoptosis, transport of bile acids and retinoid metabolism and should be considered as candidates for further mechanistic and clinical investigations.

Proteomic identification of aldo-keto reductase AKR1B10 induction after treatment of colorectal cancer cells with the proteasome inhibitor bortezomib

“…Furthermore, this enzyme is a regulator of fatty acid biosynthesis, an essential cell membrane component in cancer cells (42). Recently, the involvement of AKR1B10 has even been shown for non-small cell lung carcinoma, esophageal carcinogenesis, hepatocellular carcinoma, and uterine cancer in smokers (18,(43)(44)(45). Because of our observation of AKR1B10 induction following therapy with bortezomib and its reported biological functions, this protein was chosen for further functional studies.…”

“…These standard gels were then matched to yield information about differentially expressed proteins. We selected spots overexpressed or underexpressed by more than 4-fold to exclude artifacts caused by gel-to-gel variation observed with silver staining, and then checked spots by eye in order to avoid possible software errors (13,18).…”

Laboratory predictors of gastrointestinal tract diseases

“…За-болевание характеризуется риском развития жизне-угрожающих осложнений, таких как пищевод Бар-ретта и аденокарцинома пищевода. На сегодняшний день в литературе имеются данные о единичных протеомных исследованиях при гастроэзофагеаль-ной рефлюксной болезни [8][9][10][11]. J. Breton и соавт.…”

“…J. Breton и соавт. уделили особое внимание протеомному скринингу клеток линейной модели канцерогенеза пищево-да [9]. Согласно результатам проведенного иссле-дования, экспрессия регулятора апоптоза катепсина D и альдо-кеторедуктазы 1В10 повышена в клеточ-ных линиях, подверженных метаплазии и диспла-зии, с последовательным увеличением уровня этих протеинов в зависимости от степени трансдиффе-ренцировки поражения и, наоборот, снижением уровня указанных протеинов при аденокарциноме пищевода.…”

Lipid Peroxidation in Colorectal Carcinogenesis: Bad and Good News

“…The isoform AKR1B10 seems to be particulary involved in the transformation of HNE to the oxidized counterpart 4-oxonon-2-enal (4-ONE) (Martin et al, 2009). AKR1B10 is also up-regulated in many types of solid tumors (Fukumoto et al, 2005;Yoshitake et al, 2007;Breton et al, 2008;Satow et al, 2010), and its gene silencing results in growth inhibition of colorectal cancer cells (Yan et al, 2007), as well as in increasing HNE-elicited cell death (Matsunaga et al, 2011). Recently, some family members of AKR enzymes have been shown to be overexpressed and linked to resistance against anticancer drugs such as anthracyclines, cisplatin, and methotrexate (Veitch et al, 2009;Cheng al., 2008;Selga et al, 2008).…”