Comprehensive Characterization of Annexin I Alterations in Esophageal Squamous Cell Carcinoma

Abstract

Purpose:The purpose is to characterize alterations of the annexin I gene, its mRNA, and protein expression in esophageal squamous cell carcinoma.Experimental Design: Fifty-six cases of esophageal squamous cell carcinoma were analyzed using four microsatellite markers flanking the annexin I gene (9q11-q21) to identify loss of heterozygosity. In addition, we performed (a) single-strand conformation polymorphism and DNA sequencing along the entire promoter sequence and coding region to identify mutations, (b) real-time quantitative reverse transcription-PCR of RNA from frozen esophageal squamous cell carcinoma tissue (n ‫؍‬ 37) and in situ hybridization (n ‫؍‬ 5) on selected cases to assess mRNA expression, and (c) immunohistochemistry (n ‫؍‬ 44) to evaluate protein expression. The prevalence of the allelic variants identified in the first 56 patients was refined in 80 additional esophageal squamous cell carcinoma patients and 232 healthy individuals.Results: Forty-six of 56 (82%) esophageal squamous cell carcinoma patients showed loss of an allele at one or more of the four microsatellite markers; however, only one (silent) mutation was seen. Two intragenic variants were identified with high frequency of allelic loss (A58G, 64%; L109L, 69%). Thirty of 37 (81%) esophageal squamous cell carcinoma patients showed reduced annexin I mRNA expression, which was confirmed by in situ hybridization, whereas annexin I protein expression was reduced in 79% of poorly differentiated tumor cell foci but in only 5% of well-differentiated tumor foci, although allelic loss on chromosome 9 was found in both tumor grades.Conclusions: Allelic loss of annexin I occurs frequently, whereas somatic mutations are rare, suggesting that annexin I is not inactivated in esophageal squamous cell carcinoma via a two-hit mechanism. A decrease in annexin I protein expression was confirmed, consistent with a quantitative decrease in mRNA expression, and appeared to be related to tumor cell differentiation. We conclude that annexin I is not the tumor suppressor gene corresponding to the high levels of loss of heterozygosity observed on chromosome 9 in esophageal squamous cell carcinoma; however, dysregulation of mRNA and protein levels is associated with this tumor type.

Characterisation and protein expression profiling of annexins in colorectal cancer

“…Loss of annexin A1 has been found to be an early event in oesophageal squamous cell carcinoma, and it may function as a tumour suppressor in the development of this type of tumour (Paweletz et al, 2000). Expression of annexin A1 has also been associated with poorly differentiated oesophageal tumours of higher tumour stage (Hu et al, 2004). Together, these studies suggest that dysregulation of annexin A1 loss is important in oesophageal tumour development and progression.…”

“…In support of these findings, several studies have implicated individual annexins in tumorigenesis. Annexin A1 has been reported to show altered expression in a variety of different cancers, including oesophageal cancer (Paweletz et al, 2000;Hu et al, 2004;Wang et al, 2006), pancreatic cancer (Bai et al, 2004) and hairy cell leukaemia (Falini et al, 2004). In oesophageal adenocarcinoma, the tumour cell expression of annexin A1 has been associated with poor prognosis (Wang et al, 2006).…”

MicroRNA-196a targets annexin A1: a microRNA-mediated mechanism of annexin A1 downregulation in cancers

“…In esophageal cancers, several published reports confirm that the loss of ANXA1 is an early event in tumorigenesis (Paweletz et al, 2000;Xia et al, 2002;Hu et al, 2004). Western blotting and immunohistochemical analysis showed a drastic decrease or complete loss of ANXA1 levels in esophageal tumors and high-grade dysplasia when compared to the paired normal epithelium, implying that ANXA1 plays a functional role in maintaining the normal status of esophageal epithelium (Paweletz et al, 2000).…”

“…Suppression or loss of ANXA1 expression reported in different cancers such as prostate (Kang et al, 2002), breast (Shen et al, 2006), esophageal cancers (Paweletz et al, 2000;Xia et al, 2002;Hu et al, 2004) and B-cell non-Hodgkin's lymphoma (Vishwanatha et al, 2004) attributes tumor suppressive function to ANXA1 in these cancers. Furthermore, ANXA1 overexpression promoted caspase-mediated apoptosis in bronchoalveolar cells (Debret et al, 2003) and macrophages (Solito et al, 2001) and facilitated H 2 O 2 -induced apoptosis in thymocytes (Sakamoto et al, 1996).…”

Utility of Serum Peptidome Patterns of Esophageal Squamous Cell Carcinoma Patients for Comprehensive Treatment

“…Research showed that, if can be in the organization or the blood to the tumor markers for early stage esophageal cancer will help to make the right diagnosis, since it is difficult to get early esophageal cancer histological specimens, looking for serum markers for early diagnosis, staging and pathologic types of distinguish has important significance. In the past ten years, have found some of esophageal cancer markers, such as carcinoembryonic antigen (CEA), serum amyloid A (SAA), calcium phospholipid binding protein I (Annexin I) and glutamine transferase 3 (TGM 3), etc (Hu et al, 2004;Du et al, 2007;Banki et al, 2008;Uemura et al, 2009). It has been proved that some markers is able to assess the clinical treatment effect.…”