Objective. The anti-inflammatory proteins annexin-A1 and galectin-1 have been associated with tumor progression. This scenario prompted us to investigate the relationship between the gene and protein ...expression of annexin-A1 (ANXA1/AnxA1) and galectin-1 (LGALS1/Gal-1) in an inflammatory gastric lesion as chronic gastritis (CG) and gastric adenocarcinoma (GA) and its association with H. pylori infection. Methods. We analyzed 40 samples of CG, 20 of GA, and 10 of normal mucosa (C) by the quantitative real-time PCR (qPCR) technique and the immunohistochemistry assay. Results. High ANXA1 mRNA expression levels were observed in 90% (36/40) of CG cases (mean relative quantification RQ = 4.26 ± 2.03) and in 80% (16/20) of GA cases (mean RQ = 4.38 ± 4.77). However, LGALS1 mRNA levels were high (mean RQ = 2.44 ± 3.26) in 60% (12/20) of the GA cases, while low expression was found in CG (mean RQ = 0.43±3.13; P<0.01). Normal mucosa showed modest immunoreactivity in stroma but not in epithelium, while stroma and epithelium displayed an intense immunostaining in CG and GA for both proteins. Conclusion. These results have provided evidence that galectin-1 and mainly annexin-A1 are overexpressed in both gastritis and gastric cancer, suggesting a strong association of these proteins with chronic gastric inflammation and carcinogenesis.
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DOBA, FZAB, GIS, IJS, IZUM, KILJ, NLZOH, NUK, OILJ, PILJ, PNG, SAZU, SBCE, SBMB, SIK, UILJ, UKNU, UL, UM, UPUK
Objective. Annexin-A1 (ANXA1/AnxA1) and galectin-1 (LGALS1/Gal-1) are mediators that play an important role in the inflammatory response and are also associated with carcinogenesis. We investigated ...mRNA and protein expression in precancerous gastric lesions that participate in the progression cascade to gastric cancer, such as intestinal metaplasia (IM) and gastric ulcer (GU). Methods. Quantitative real-time PCR (qPCR) and immunohistochemical techniques were used to analyze the relative quantification levels (RQ) of ANXA1 and LGALS1 mRNA and protein expression, respectively. Results. Increased relative expression levels of ANXA1 were found in 100% of cases, both in IM (mean RQ = 6.22 ± 0.06 ) and in GU (mean RQ = 6.69 ± 0.10 ). However, the LGALS1 presented basal expression in both groups (IM: mean RQ = 0.35 ± 0.07 ; GU: mean RQ = 0.69 ± 0.09 ). Immunohistochemistry revealed significant positive staining for both the AnxA1 and Gal-1 proteins in the epithelial nucleus and cytoplasm as well as in the stroma of the IM and GU groups ( P < 0.05 ) but absence or low immunorectivity in normal mucosa. Conclusion. Our results bring an important contribution by evidencing that both the AnxA1 and Gal-1 anti-inflammatory proteins are deregulated in precancerous gastric lesions, suggesting their involvement in the early stages of gastric carcinogenesis, possibly due to an inflammatory process in the gastric mucosa.
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DOBA, FZAB, GIS, IJS, IZUM, KILJ, NLZOH, NUK, OILJ, PILJ, PNG, SAZU, SBCE, SBMB, SIK, UILJ, UKNU, UL, UM, UPUK
Functional polymorphisms in promoter regions can produce changes in the affinity of transcription factors, thus altering the messenger ribonucleic acid (mRNA) expression levels of inflammatory ...cytokines associated with the risk of cancer development. The goal of this study was to evaluate the influence that polymorphisms in the cytokine genes known as
TNF
-α-308 G/A (rs1800629),
TNF
-α-857 C/T (rs1799724),
IL
-
8
-251 T/A (rs4073),
IL
-
8
-845 T/C (rs2227532), and
IL
-
10
-592 C/A (rs1800872) have on changes to mRNA expression levels and on the risks of chronic gastritis (CG) and gastric cancer (GC). A sample of 723 individuals was genotyped using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique. Relative mRNA expression levels were measured using quantitative real-time PCR (qPCR). Polymorphisms
TNF
-
α
-308 G/A and
IL
-
8
-251 A/T were not associated with risks of these gastric lesions. However,
TNF
-
α
-857 C/T,
IL
-
8
-845 T/C, and
IL
-
10
-592 C/A were found to be associated with a higher risk of GC, and
IL
-
10
-592 C/A was found to be associated with a higher risk of CG. The relative mRNA expression levels (RQ) of
TNF
-
α
,
IL
-
8
, and
IL
-
10
were markedly downregulated in the CG group (median RQs = 0.128, 0.247, and 0.614, respectively), while the RQ levels of
TNF
-
α
in the GC group were upregulated (RQ = 2.749), but were basal for
IL
-
8
(RQ = 1.053) and downregulated for
IL
-
10
(RQ = 0.179). When the groups were stratified according to wild-type and polymorphic alleles, only for
IL
-
8
-845 T/C the polymorphic allele was found to influence the expression levels of this cytokine.
IL
-
8
-845 C allele carriers were significantly upregulated in both groups (GC and CG; RQ = 3.138 and 2.181, respectively) when compared to TT homozygotes (RQ = −0.407 and 0.165, respectively). In silico analysis in the
IL
-
8
promoter region revealed that the presence of the variant C allele in position -845 is responsible for the presence of the binding sites for two transcription factors (
REL
and
CREB1
), which are involved in increased gene expression. Polymorphic alleles were not shown to have any effect on the expression levels of
TNF
-
α
and
IL
-
10
. Taken together, our findings provide evidence for an association of
TNF
-
α
-857 C/T,
IL
-
8
-845 T/C, and
IL
-
10
-592 C/A with a higher risk of gastric cancer and also demonstrate the influence that the polymorphic C allele of
IL
-
8
-845 has on changes to the gene expression levels of this cytokine.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, MFDPS, NUK, OILJ, PNG, SAZU, SBCE, SBJE, SBMB, SBNM, UL, UM, UPUK, VKSCE, ZAGLJ
Functional polymorphisms in promoter regions can produce changes in the affinity of transcription factors, thus altering the messenger ribonucleic acid (mRNA) expression levels of inflammatory ...cytokines associated with the risk of cancer development. The goal of this study was to evaluate the influence that polymorphisms in the cytokine genes known as TNF-alpha-308 G/A (rs1800629), TNF-alpha-857 C/T (rs1799724), IL-8-251 T/A (rs4073), IL-8-845 T/C (rs2227532), and IL-10-592 C/A (rs1800872) have on changes to mRNA expression levels and on the risks of chronic gastritis (CG) and gastric cancer (GC). A sample of 723 individuals was genotyped using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique. Relative mRNA expression levels were measured using quantitative real-time PCR (qPCR). Polymorphisms TNF-alpha-308 G/A and IL-8-251 A/T were not associated with risks of these gastric lesions. However, TNF-alpha-857 C/T, IL-8-845 T/C, and IL-10-592 C/A were found to be associated with a higher risk of GC, and IL-10-592 C/A was found to be associated with a higher risk of CG. The relative mRNA expression levels (RQ) of TNF-alpha, IL-8, and IL-10 were markedly downregulated in the CG group (median RQs=0.128, 0.247, and 0.614, respectively), while the RQ levels of TNF-alpha in the GC group were upregulated (RQ=2.749), but were basal for IL-8 (RQ=1.053) and downregulated for IL-10 (RQ=0.179). When the groups were stratified according to wild-type and polymorphic alleles, only for IL-8-845 T/C the polymorphic allele was found to influence the expression levels of this cytokine. IL-8-845 C allele carriers were significantly upregulated in both groups (GC and CG; RQ=3.138 and 2.181, respectively) when compared to TT homozygotes (RQ=-0.407 and 0.165, respectively). In silico analysis in the IL-8 promoter region revealed that the presence of the variant C allele in position -845 is responsible for the presence of the binding sites for two transcription factors (REL and CREB1), which are involved in increased gene expression. Polymorphic alleles were not shown to have any effect on the expression levels of TNF-alpha and IL-10. Taken together, our findings provide evidence for an association of TNF-alpha-857 C/T, IL-8-845 T/C, and IL-10-592 C/A with a higher risk of gastric cancer and also demonstrate the influence that the polymorphic C allele of IL-8-845 has on changes to the gene expression levels of this cytokine.
Full text
Available for:
GEOZS, IJS, IMTLJ, KILJ, KISLJ, MFDPS, NUK, OILJ, PNG, SAZU, SBCE, SBJE, SBMB, SBNM, UL, UM, UPUK, VKSCE, ZAGLJ
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No presente estudo foram investigados os níveis de expressão gênica e protéica da anexina-1 (ANXA1/AnxA1) e galectina-1 (LGALS1/Gal-1) na carcinogênese do estômago e associações com infecção pela Helicobacter pylori e o genótipo de virulência bacteriano cagA+. A análise foi realizada em 40 biópsias de mucosa gástrica com gastrite crônica (CG), 20 de câncer gástrico (GA) e 10 de mucosa normal (C), pelas técnicas de qPCR para quantificar os níveis de RNAm; imuno-histoquímica para caracterizar a expressão protéica na mucosa gástrica, e PCR para diagnóstico molecular da H. pylori e cepa cagA+. O estudo mostrou resultados inéditos quanto à expressão desses genes em gastrite crônica, ainda sem descrições na literatura. Foi demonstrada expressão relativa elevada do mRNA de ANXA1 em 80% dos casos de GA (média de 4,38 + 4,77) e em 90% dos casos de CG (média de 4,26 + 2,03), sem diferença significante entre os grupos (p = 0,33). O gene LGALS1 apresentou expressão elevada em 60% dos casos GA (média de 2,44 + 3,26) e, expressão constitutiva na CG (média de 0,43 + 3,13), mostrando, portanto, diferença significante entre os grupos (p < 0,01). A imuno-histoquímica revelou que as proteínas AnxA1 e Gal-1 não são expressas na mucosa normal. Ao contrário, durante o processo inflamatório de CG, imunomarcação citoplasmática positiva para a AnxA1 foi observada na porção basal do epitélio e estroma e, para Gal-1 a expressão foi constatada na porção apical e borda estriada do epitélio além do estroma. No adenocarcinoma tipo intestinal foi observada expressão citoplasmática em toda extensão epitelial e estroma tanto para a AnxA1 quanto para a Gal-1. Por outro lado, no tipo difuso imunomarcação positiva também foi observada no núcleo e membrana plasmática...
In this study we investigated the levels of gene and protein expression of annexin-1 (ANXA1/Anxa1) and galectin-1 (LGALS1/Gal-1) in gastric carcinogenesis and associations with Helicobacter pylori infection and bacterial virulence genotype cagA+. The analysis was performed in 40 biopsies of gastric mucosa with chronic gastritis (CG), 20 with gastric cancer (GA) and 10 of normal mucosa (C), by the techniques of qPCR to quantify mRNA levels, immunohistochemistry to characterize the protein expression in gastric mucosa, and PCR for molecular diagnosis of H. pylori cagA+ strains. This is the first study regarding the expression of these genes in chronic gastritis. High ANXA1expression levels were demonstrated in 80% of GA cases (mean 4.38 + 4.77) and in 90% of GC cases (mean 4.26 + 2.03), with no significant difference between groups (p = 0.33). High LGALS1 gene expression was found in 60% of GA cases (average 2.44 + 3.26), and constitutive expression was found in CG (mean 0.43 + 3.13), showing therefore a significant difference between groups (p <0.01). Immunohistochemistry revealed that the proteins AnxA1 and Gal-1 are not expressed in normal mucosa. In contrast, during the inflammatory process of CG, positive cytoplasmic immunostaining for AnxA1 was observed in the basal epithelium and stroma, and Gal-1 expression was detected in the apical portion and striated border of the epithelium and stroma. In intestinal-type adenocarcinoma was observed cytoplasmic expression in all epithelial and stromal extension for both AnxA1 and Gal-1. On the other hand, in diffuse-type adenocarcinoma positive immunostaining was also observed in the nucleus and plasma membrane of both proteins. Infection by H. pylori showed no association with the expression of both genes, but the genotype cagA+ is associated with about 2.5 times... (Complete abstract click electronic access below)
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