Márcia Valéria Pitombeira Ferreira

The relationship between Helicobacter pylori genes cagE and virB11 and gastric cancer

BACKGROUND The association between Helicobacter pylori gene diversity and gastric cancer has been poorly reported, although it is one of the important ways to explain the gastric pathogenesis. The aim of this study was to investigate the frequency of cagE and virB11 genes in H. pylori isolated from patients with gastric cancer and to analyze the histology profiles. MATERIALS AND METHODS The presence of H. pylori and subtypes (cagE and virB11) was detected by PCR from the genomic DNA of 101 patients who had been diagnosed with gastric cancer. The cases were grouped according to the presence/absence of the genes studied and were analyzed in relation to histopathological parameters. RESULTS H. pylori infection was detected in 94 out of 101 (93.1%) gastric carcinomas. The cases were categorized into the following groups: cagE+/virB11+, cagE+/virB11-, cagE-/virB11+, and cagE-/virB11-. Frequencies were: 50% (47/94) cagE+/virB11+, 3.2% (3/94) cagE+/virB11-, 10.6% (10/94) cagE-/virB11+, and 36.2% (34/94) cagE-/virB11-. Tumors in the gastric antrum were predominant. An exception was the cagE-/virB11- group, in which tumors had a tendency to be located in the gastric cardia; the majority of the cardia tumors (56% (14/25)) were in this group. Intestinal histology type was the most frequent, but the cagE+/virB11- group only had diffuse tumors. H. pyloricagE+/virB11+ occurred most frequently (except at stage III), and was present at all gastric cancer stages. CONCLUSIONS This study is the first to include a relevant number of gastric cancer cases with H. pylori infection, reporting the frequency and relationship of cagE and virB11 genes and the genesis of this tumor. The presence of these cag pathogenicity island genes shows that they are important factors for the pathogenesis and malignancy of gastric cancer related to H. pylori.

Inactivation of COX-2, HMLH1 and CDKN2A Gene by Promoter Methylation in Gastric Cancer: Relationship with Histological Subtype, Tumor Location and Helicobacter pylori Genotype

Objective: We aimed to evaluate the inactivation of COX-2, HMLH1 and CDKN2A by promoter methylation and its relationship with the infection by different Helicobacter pylori strains in gastric cancer. Methods: DNA extracted from 76 H. pylori-positive gastric tumor samples was available for promoter methylation identification by methylation-specific PCR and H. pylori subtyping by PCR. Immunohistochemistry was used to determine COX-2, p16INK4A and HMLH1 expression. Results: A strong negative correlation was found between the expression of these markers and the presence of promoter methylation in their genes. Among cardia tumors, negativity of p16INK4A was a significant finding. On the other hand, in noncardia tumors, the histological subtypes had different gene expression patterns. In the intestinal subtype, a significant finding was HMLH1 inactivation by methylation, while in the diffuse subtype, CDKN2A inactivation by methylation was the significant finding. Tumors with methylated COX-2 and HMLH1 genes were associated with H. pylori vacA s1 (p = 0.025 and 0.047, respectively), and the nonmethylated tumors were associated with the presence of the gene flaA. Conclusions: These data suggest that the inactivation of these genes by methylation occurs by distinct pathways according to the histological subtype and tumor location and depends on the H. pylori genotype.

Effects of Amburoside A and Isokaempferide, Polyphenols from Amburana cearensis, on Rodent Inflammatory Processes and Myeloperoxidase Activity in Human Neutrophils

The present study evaluated the anti-inflammatory activity of amburoside A (a phenol glucoside) and isokaempferide (a flavonol) isolated from the trunk bark of Amburana cearensis, a medicinal plant used in northeast Brazil for the treatment of asthma. Animals (male Wistar rats or Swiss mice) pre-treated with amburoside A (25 and 50 mg/kg) or isokaempferide (12.5, 25 and 50 mg/kg), orally or intraperitoneally, showed a significant inhibition of the paw oedema induced by carrageenan (1%), prostaglandin E(2) (30 nmol/paw), histamine (200 microg/paw) or serotonin (200 microg/paw). Histological and morphometric evaluations of the rat paw oedema induced by carrageenan showed that amburoside A and isokaempferide also inhibited the accumulation of inflammatory cells. Amburoside A reduced significantly the paw oedema and the increase in vascular permeability induced by dextran, as related to the control group. Similar results were observed with the isokaempferide pre-treatment. Furthermore, amburoside A or isokaempferide inhibited both leucocyte and neutrophil migrations, in mouse peritoneal cavity, after the carrageenan injection. The polyphenols were not cytotoxic and blocked N-formyl-methyl-leucyl-phenylalanine-induced myeloperoxidase release and activity in human neutrophils. In addition, amburoside A and isokaempferide at 50 and 100 microg/ml concentrations reduced significantly the lipopolysaccharide-mediated increase in tumour necrosis factor-alpha (TNF-alpha) levels. These results provide, for the first time, evidence to support the anti-inflammatory activity of amburoside A and isokaempferide that seems to be related to an inhibition of inflammatory mediators, such as TNF-alpha, as well as histamine, serotonin and prostaglandin E(2), besides leucocyte infiltration in a dose- or concentration-dependent manner. These anti-inflammatory effects can be explained, at least in part, by the ability of these compounds to reduce neutrophil degranulation, myeloperoxidase activity, mediators as well as TNF-alpha secretion.

Gastric adenocarcinoma and Helicobacter pylori: correlation with p53 mutation and p27 immunoexpression.

INTRODUCTION Helicobacter pylori infection is an established risk factor for gastric cancer development, but the exact underlying mechanism still remains obscure. The inactivation of tumor suppressor genes such as p53 and p27(KIP1) is a hypothesized mechanism, although there is no consensus regarding the influence of H. pylori cagA(+) in the development of these genetic alterations. GOALS To verify the relationship among H. pylori infection, p53 mutations and p27(Kip1) Protein (p27) expression in gastric adenocarcinomas (GA) seventy-four tissues were assayed by PCR for H. pylori and cagA presence. Mutational analysis of p53 gene was performed by single-strand conformation polymorphism (SSCP). Seventy tissues were analyzed by an immunohistochemical method for p27 expression. RESULTS From the samples examined, 95% (70/74) were H. pylori positive, 63% cagA(+). Altered p53 electrophoretic mobility was found in 72% of cases and significantly more frequent in the presence of cagA. Considerable reduction in p27 expression (19%) was found with a tendency for association between cagA(+) and p27(-), although the results were not statistically significant. Concomitant alterations of both suppressor genes were detected in 60% of cases. In the cases cagA(+), 66.7% of them had these concomitant alterations. CONCLUSIONS The data suggest that H. pylori cagA(+) contributes to p53 alteration and indicate that concomitant gene inactivation, with reduced p27 expression, may be a mechanism in which H. pylori can promote the development and progression of gastric cancer.

Helicobacter pylori genotype and polymorphisms in DNA repair enzymes: Where do they correlate in gastric cancer?

One of the mechanisms proposed by which H. pylori causes gastric cancer (GC) is through DNA damage due to chronic inflammation. Genomic integrity is guaranteed by repair enzymes such as APE‐1, OGG‐1, and PARP‐1. Host genetic polymorphisms associated with the bacterial strain may influence the ability to repair the damage, contributing to the development of H. pylori‐associated GC. The aim of this study was to determine the association of the polymorphisms APE‐1 (T2197G), OGG‐1 (C1245G), and PARP‐1 (A40676G) with H. pylori‐genotype in 109 patients with GC.

Neuropathology of two Brazilian autopsied cases of tropical spastic paraparesis / HTLV-I associated myelopathy (TSP/HAM) of long evolution.

We report on a neuropathological analysis of two cases of TSP/HAM originating from Brazil. These two cases had, respectively, an evolution of 13 and 40 years. The main neuropathological findings consisted of spinal cord atrophy, mainly the lower thoracic cord, diffuse degeneration of the white and grey matter, rare foci of mononuclear and perivascular cuffs, and hyaline hardening of arteriolae. The supraspinal structures were normal, excepting for a slight gliosis in the cerebellum. An analysis on the long evolutive cases as described in the literature is outlined in this study.

p27KIP1 expression in gastric cancer: differential pathways in the histological subtypes associated with Helicobacter pylori infection.

Abstract Objective. Decreases in p27KIP1 and C-MYC expression have been associated with Helicobacter pylori infection. Furthermore, C-MYC seems to be a transcriptional repressor of p27KIP1. Therefore, in a series of gastric adenocarcinomas we studied the association of p27KIP1 expression with H. pylori genotype (vacA, cagA, cagE and virB11) and the involvement of C-MYC in this process. Material and methods. Expression of p27KIP1 and C-MYC was determined by immunohistochemistry in 84 gastric adenocarcinoma samples and H. pylori infection and genotype were determined by polymerase chain reaction. Results. Most p27KIP1-negative cases (94.0%) were H. pylori-positive and 44.8% were C-MYC-positive. In the diffuse gastric cancer subtype, p27-negative-C-MYC-positive was the most frequent combination (cluster II), and was associated with the more pathogenic H. pylori strains. Although an association with p27KIP1 and H. pylori strain was found in the intestinal gastric cancer subtype, negativity for p27KIP1 and C-MYC markers was the most frequent cluster, followed by cluster II, and both were present, independent of the H. pylori genotype. Conclusions. Reduced expression of p27KIP1 was closely linked to H. pylori infection, and was dependent on the more pathogenic strains. Moreover, intestinal and diffuse subtypes showed distinct carcinogenic pathways influenced by H. pylori strains. These data add insight to the differential influence and relevance of H. pylori genotype in gastric cancer development.

Relationship between EBV infection and expression of cellular proteins c-Myc, Bcl-2, and Bax in gastric carcinomas.

Background Epstein-Barr virus (EBV) has been related to tumorigenesis in about 10% of all gastric carcinomas. Several studies have demonstrated strong evidence of its involvement in this process, but most of the mechanisms used by the virus to control this process are still unknown. Previous studies in vitro have indicated a relationship between the virus and some cellular genes involved in processes such as proliferation and apoptosis. Objective The aim of the present study was to investigate a possible EBV-induced tumorigenic pathway involving the cellular proteins Bcl-2, Bax, and c-Myc. Study Design One hundred patients of gastric carcinoma, obtained from 2 hospitals in Fortaleza, Brazil were assessed for the presence of EBV by in situ hybridization, for the expression of Bcl-2, Bax, and c-Myc (nuclear and cytoplasmic staining) proteins by immunohistochemistry techniques, and for the apoptotic index. Results EBV was detected in 8 (8%) patients showing strong staining situated in the nuclei of the tumor cells, 6 of them displaying a diffuse pattern, and 2 demonstrating a focal pattern of staining. The correlation with the immunohistochemistry results demonstrated that none of the EBV-positive cases exhibited Bcl-2 staining. On the other hand, Bax and c-Myc (nuclear) proteins demonstrated a significant positivity index and staining scores (labeling index and H-score) in the EBV-positive group; however, the values were lower than those obtained in the EBV-negative group, notably for c-Myc nuclear protein. In contrast, the cytoplasmic staining of c-Myc protein revealed slightly higher staining values in the EBV-positive group. The balance between Bcl-2 and Bax proteins demonstrated that the majority of the evaluated cases exhibited apoptosis orientation; however, in 62.5% of the EBV-positive cases neither protein was observed. The average apoptotic index was 4.58%, demonstrating a slightly lower average in the EBV-positive group. Conclusions EBV is not related to the overexpression of Bcl-2 and c-Myc (nuclear) in gastric carcinomas; however, the results point to a possible EBV involvement with the transport mechanisms of the nuclear membrane, resulting in cytoplasmic c-Myc accumulation. The suppression of Bax expression could represent an alternative viral mechanism for inhibition of apoptosis.

Characterization of Gastric Cardia Tumors: Differences in Helicobacter pylori Strains and Genetic Polymorphisms

AbstractBackgroundGastric cancer results from a multifactorial process and is one of the most common causes of death worldwide. These tumors can arise in the distal stomach (non-cardia) and in the cardia region, presenting different characteristics and frequency of occurrence worldwide.Aims To search for differences between tumors of different locations that could explain the presence of cardia tumors, considering Helicobacter pylori strains and genetic polymorphisms.Materials and MethodsDNA was extracted from gastric adenocarcinoma tissue of 127 patients. Helicobacter pylori genes were detected by PCR, and polymorphisms by PCR–RFLP.ResultsMost of the tumors were located in non-cardia. The genotype 28152GA of XRCC1 showed an increase in risk of cardia tumors. In analysis performed considering gender, women carrying TNF-308GA genotype showed a decreased risk of non-cardia tumors, while in men the decreased risk of non-cardia tumors was associated with TNF-308GG genotype. Genotypes combinations showed that the SNPs RAD51 135G>C, XRCC3 18067C>T, and XRCC1 28152G>A had some combinations more frequent in cardia tumors, with an increased risk. Patients infected by cagE-positive strains presented a positive correlation with non-cardia tumors.ConclusionThe results showed some susceptibility differences between tumors of different locations. There was an increased risk relationship between three repair enzyme SNPs and cardia tumors, and the G allele of the cytokine gene TNF negatively influenced the development of non-cardia tumors. Helicobacter pylori strains seemed to be different in the cardia region, where they were less virulent than those located in the distal region of the stomach.

Differential expression of MYC in H. pylori- related intestinal and diffuse gastric tumors

Evidence suggests that the carcinogenic process guided by Helicobacter pylori is related to the expression of cell cycle and apoptosis proteins as BCL-2, BAX, and MYC. However, the literature is conflicting regarding the expression frequency in the histological subtypes and did not consider cagA gene presence. To investigate the expression of these proteins considering the histological subtypes of gastric cancer associated with H. pylori (cagA), a total of 89 cases were used. H. pylori infection and cagA status were determined by PCR. Immunodetection was performed for MYC, BCL-2, and BAX proteins. H. pylori was found in 95.5% of the patients, among them, 65.8% were cagA(+). Nuclear MYC was detected in 36.4%, BAX in 55.7%, while BCl-2 in just 5%. Nuclear MYC staining was significantly lower in the intestinal than diffuse subtype (p = 0.008) and was related with the presence of H. pylori cagA(+). Additionally, most of the few cases cytoplasmic MYC positive were in the intestinal subtype. In diffuse tumors, although most nuclear MYC positive cases were cagA(+), it was not significant. No difference was observed between BCL-2 or BAX expression considering the presence of cagA gene in the histological subtypes. It seems that MYC could be relevant for the diffuse tumorigenic pathway associated with H. pylori and possibly influenced by the presence of cagA gene, while in intestinal tumors, the tumorigenic pathway does not occur through the MYC expression.