Cristiane Oliveira

Polymorphisms in the 5′- and 3′-untranslated region of the VEGF gene and sporadic breast cancer risk and clinicopathologic characteristics

AbstractThe wild and the variant alleles of the C936T and G634C vascular endothelial grow factor (VEGF) polymorphisms seem to be linked to higher angiogenic phenotype than the remaining alleles and may act on breast cancer (BC) origin. We investigated the influence of the VEGF C936T and G634C polymorphisms on the occurrence and clinicopathologic characteristics of sporadic breast cancer (SBC) in 235 patients and 235 controls. Peripheral blood samples of all individuals were analysed by the polymerase chain reaction for identification of genotypes and by enzyme-linked immunosorbent assay (ELISA) for quantification of serum VEGF levels. The variant 634CC genotype isolated (16.2% versus 10.7%, P  = 0.01) and in combination with the wild 936CC genotype (10.6% versus 5.5%, P  = 0.01) were more common in patients than in controls. The carriers of the respective genotypes were under a 2.20-fold and a 3.08-fold increased risks for the disease. Additionally, the frequency of the wild 936CC genotype was higher in patients with tumours of histological grade III compared to those with tumours of I+II histological grades (84.0% versus 64.7%, P  = 0.004) and in patients with positive oestrogen receptor tumours compared to those with tumours lacking oestrogen receptor expression (84.7% versus 73.9%, P  = 0.02). Similar serum values of VEGF were seen in patients and controls with the distinct genotypes of the VEGF. The data suggest that the VEGF wild 936CC and the variant 634CC genotypes constitute inherited determinants of SBC and SBC aggressiveness in Brazil, but are not significant predictors of circulating VEGF levels.

Surveillance of active human cytomegalovirus infection in hematopoietic stem cell transplantation (HLA sibling identical donor): search for optimal cutoff value by real-time PCR

BackgroundHuman cytomegalovirus (CMV) infection still causes significant morbidity and mortality after allogeneic hematopoietic stem cell transplantation (HSCT). Therefore, it is extremely important to diagnosis and monitor active CMV infection in HSCT patients, defining the CMV DNA levels of virus replication that warrant intervention with antiviral agents in order to accurately prevent CMV disease and further related complications.MethodsDuring the first 150 days after allogeneic HSTC, thirty patients were monitored weekly for active CMV infection by pp65 antigenemia, nested-PCR and real-time PCR assays. Receiver operating characteristic (ROC) plot analysis was performed to determine a threshold value of the CMV DNA load by real-time PCR.ResultsUsing ROC curves, the optimal cutoff value by real-time PCR was 418.4 copies/104 PBL (sensitivity, 71.4%; specificity, 89.7%). Twenty seven (90%) of the 30 analyzed patients had active CMV infection and two (6.7%) developed CMV disease. Eleven (40.7%) of these 27 patients had acute GVHD, 18 (66.7%) had opportunistic infection, 5 (18.5%) had chronic rejection and 11 (40.7%) died - one died of CMV disease associated with GVHD and bacterial infection.ConclusionsThe low incidence of CMV disease in HSCT recipients in our study attests to the efficacy of CMV surveillance based on clinical routine assay. The quantification of CMV DNA load using real-time PCR appears to be applicable to the clinical practice and an optimal cutoff value for guiding timely preemptive therapy should be clinically validated in future studies.

Polymorphisms of glutathione S-transferase Mu 1 (GSTM1), Theta 1 (GSTT1), and Pi 1 (GSTP1) genes and epithelial ovarian cancer risk

Background: Exposure of ovarian cells to estrogen, which is detoxified by glutathione S-transferases (GSTs), has been associated with epithelial ovarian cancer (EOC) development. Objectives: We tested in this study whether the GSTM1, GSTT1 and GSTP1 Ile105Val polymorphisms alter the risk of EOC. Materials and methods: Genomic DNA from 132 EOC patients and 132 controls was analyzed by polymerase chain reaction and restriction fragment length polymorphism methods. The differences between groups were analyzed by χ2 or Fisher’s exact test. Results: The frequencies of GSTP1 Ile/Ile (57.6% versus 45.5%, P = 0.03), GSTM1 null plus GSTP1 Ile/Ile (43.5% versus 25.8%; P = 0.03) and GSTM1 null plus GSTT1 null plus GSTP1 Ile/Ile (30.3% versus 7.7%; P = 0.007) genotypes were higher in patients than in controls. Individuals with the respective genotypes had a 1.80 (95% CI: 1.06–3.06), 2.38 (95% CI: 1.08–5.24) and 11.28 (95%CI: 1.95–65.30)-fold increased risks of EOC than those with the remaining genotypes. Conclusions: Our data present preliminary evidence that GSTM1, GSTT1 and GSTP1 polymorphisms, particularly in combination, constitute important inherited EOC determinants in individuals from Southeastern Brazil.

Biochemical imaging of normal, adenoma, and colorectal adenocarcinoma tissues by Fourier transform infrared spectroscopy (FTIR) and morphological correlation by histopathological analysis: preliminary results

Introduction: The colorectal cancer is a major health problem worldwide. Histology is considered the gold standard for differential diagnosis. However, it depends on the observers experience, which can lead to discrepancies and poor results. Spectroscopic imaging by Fourier transform infrared (FTIR) is a technique that may be able to improve the diagnosis, because it is based on biochemical differences of the structural constituents of tissue. Therefore, the main goal of this study was to explore the use of FTIR imaging technique in normal colon tissue, colorectal adenoma, and adenocarcinoma in order to correlate their morphological structures with their biochemical imaging. Methods: Samples were collected from normal (n = 4), adenoma (n = 4), and adenocarcinoma human colorectal tissue (n = 4) from patients undergoing colonoscopy or surgical resection of colon lesions. The samples were sectioned with a cryostat in sequential sections; the first slice was placed on CaF 2 slide and the second slice was placed on glass slide for histological analysis (HE staining). The cluster analyses were performed by the software Cytospec (1.4.02)  . Results: In normal samples, biochemical analysis classified six different structures, namely the lamina propria of mucous glands (epithelial cells and goblet cells), central lumen of the gland, mucin, and conjunctive tissue. In samples with adenoma and adenocarcinoma, altered regions could also be identified with high sensitivity and specificity. Conclusion: The results of this study demonstrate the potential and viability of using infrared spectroscopy to identify and classify colorectal tissues.

Association between genetic polymorphisms in apoptosis-related genes and risk of cutaneous melanoma in women and men.

BACKGROUND The P53 Arg72Pro, MDM2 c.+309T>G, BAX c.-248G>A, and BCL2 c.-717C>A polymorphisms have variable roles in the apoptosis pathways. OBJECTIVE To clarify the roles of these polymorphisms in the risk for cutaneous melanoma (CM). METHODS Genomic DNA of 200 CM patients and 215 controls was analyzed by PCR-RFLP. RESULTS In women, the frequencies of BAX GG (83.0% vs. 71.0%, P=0.04), BCL2 AA (32.0% vs. 15.0%, P=0.003), P53 ArgArg plus BAX GG (84.9% vs. 63.2%, P=0.01), P53 ArgArg plus BCL2 AA (37.0% vs. 13.1%, P=0.003), BAX GG plus BCL2 AA (70.3% vs. 33.3%, P=0.001), MDM2 GG plus BAX GG plus BCL2 AA (27.3% vs. 3.7%, P=0.03), and P53 ArgArg plus MDM2 GG plus BAX GG plus BCL2 AA (33.3% vs. 5.6%, P=0.04) genotypes were higher in patients than in controls. Female carriers of the respective genotypes were under 1.98 (95% CI: 1.01-3.91), 2.87 (95% CI: 1.43-5.77), 3.48 (95% CI: 1.34-9.04), 4.23 (95% CI: 1.63-10.96), 6.04 (95% CI: 2.10-17.37), 25.61 (95% CI: 1.29-507.24), and 25.69 (95% CI: 1.11-593.59)-fold increased risks for CM than others, respectively. In men, the frequencies of BCL2 CA+AA (83.0% vs. 67.6%, P=0.01) and MDM2 TG+GG plus BCL2 CA+AA (94.2% vs. 68.3%, P=0.003) genotypes were higher in patients than in controls. Male carriers of the respective genotypes were under 2.43 (95% CI: 1.23-4.82) and 9.22 (95% CI: 2.16-39.31)-fold increased CM risks than others, respectively. CONCLUSION The data suggest for the first time that P53 Arg72Pro, MDM2 c.+309T>G, BAX c.-248G>A, and BCL2 c.-717C>A polymorphisms, enrolled in apoptosis pathways, constitute distinct determinants of CM in women and men.

PDCD1 Gene Polymorphisms as Regulators of T‐Lymphocyte Activity in Cutaneous Melanoma Risk and Prognosis

This study aimed to evaluate whether PD1.1 (c.‐606G>A), PD1 (c.627 + 252C>T), PD1.5 (c.804C>T), and PD1.9 (c.644C>T) single nucleotide polymorphisms of PDCD1 gene influence the risk, clinicopathological aspects, and survival of cutaneous melanoma (CM). Individuals with phototype I or II and PD1 CC genotype were under 5.89‐fold increased risk of developing CM. PD1.5 TT genotype increased PDCD1 expression (2.49 versus 1.28 arbitrary units, p = .03) and PD1.5 CT or TT genotype and allele T increased PD1 expression in TCD4+ lymphocytes (16.6 versus 12.5%, p = .01; 17.0 versus 13.1%, p = .006). At 60 months of follow‐up, short recurrence‐free survival was seen in patients with PD1.1 AA genotype (33.3 versus 71.8%, p = .03). Patients with PD1.1 AA and PD1.5 CC genotype had 4.21 and 2.62 more chances of presenting relapse and evolving death by disease in Cox analyses, respectively. Our data provide preliminary evidence that abnormalities in regulation of T lymphocyte alter CM risk, clinical aspects, and prognosis.

Polymorphisms in key regulator genes of the intrinsic apoptosis pathway in risk and clinical presentation of diffuse large B-cell lymphoma

To the Editor Diffuse large B-cell lymphoma (DLBCL) is the most common subtype of non-Hodgkin lymphoma (NHL), accounting for 30–40% of the newly diagnosed cases. Apoptosis is an important mechanism involved in lymphomagenesis, in which the pro-apoptotic B-cell lymphoma 2-associated X (Bax) and the anti-apoptotic Bcell lymphoma 2 (Bcl2) are the most active proteins [1]. It is already well established that the ability to induce apoptosis is variable in humans [2]. A single nucleotide polymorphism (SNP) located in the 5′-untranslated region of the BAX gene with a G→A substitution at 248 nucleotide position (rs4645878) has its allele ‘G’ associated with lower transcriptional activity when compared with ‘A’ allele [3]. Regarding BCL2 gene, a C→A substitution at 717 nucleotide position (rs2279115), located on the promoter region of the gene, has its AA genotype associated with increased Bcl2 expression in comparison with CC genotype [4]. Wang et al. [5] found excesses of the GA+AA genotype of BAX c. 248G>A and the AA genotype of BCL2 c.938C>A (rs2279115) SNPs in a mixed group of NHL patients compared to controls in China. The authors also found that the above mentioned genotypes were associated with dimension and stage of the tumour, respectively. Because DLBCL has a distinct behaviour in comparison with other types of NHL [6], and genetic variations in genes related to cellular homeostasis might have a crucial role in DLBCL development [7], the identification of BAX c. 248G>A and BCL2 c. 717C>A genotypes in patients with de novo DLBCL and controls was considered necessary to test whether SNPs influence the risk and clinical presentation of this aggressive NHL. The case group comprised 153 de novo DLBCL patients at diagnosis (median age: 56 years, range: 17–89 years, 70 males, 83 females, 140 Caucasians, 13 non-Caucasians), considering that 101 patients presented systemic symptoms seen at the Haematology and Haemotherapy Centre from December 2007 to June 2014. The diagnosis of DLBCL was established in accord with the World Health Organization criteria. R-IPI was calculated in each case as previously reported [8], and 16 patients were of very good R-IPI, 76 of good R-IPI, and 61 of poor R-IPI. The control group comprised 240 blood donors (median age: 49 years, range: 23–60 years, 124 males, 116 females,

Association of BAX G(-248)A and BCL2 C(-717)A polymorphisms with outcome in diffuse large B-cell lymphoma patients.

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278PASSOCIATION OF ERP29 GENETIC POLYMORPHISM WITH BREAST CANCER RISK AND PROGNOSIS

ABSTRACT Aim: ERP29, a tumor suppressor gene, was related with the onset of tumors. The influence of ERP29 c.*293A > G (rs7114) polymorphism in breast cancer (BC) risk and prognosis has never been performed before and was the aim of this study. Methods: We analyzed 737 BC patients and 742 healthy women. The BC patients were treated by conventional procedures. ERP29 c.*293A > G genotypes were obtained from genomic DNA by TaqMan® genotyping assays. The expression of ERP29 mRNA was determined by quantitative PCR using total RNA from blood leukocytes of healthy individuals with distinct genotypes (15 individuals with AA, 17 with AG, and six with GG). Overall survival (OS) was obtained from date of first diagnosis until the date of death or last follow-up. The differences in frequencies of distinct genotypes in patients and controls were analyzed by logistic regression model, serving to obtain age, ethnic origin and adjusted crude odds ratios (ORs), considering a confidence interval (CI) of 95%. OS time was calculated using the Kaplan-Meier estimate probabilities and differences between survival curves were analyzed by the log-rank test. Statistical significance was established at a P Results: The frequency of ERP29 AG + GG combined genotypes was higher in BC patients than in controls (36.5% vs. 30.7%, P= 0.03). Carriers of G allele were under a 1.32 (95% CI: 1.02-1.70)-fold increased risk for the tumor. The mRNA expression was lower in carriers of G allele than those with ERP29 AA genotype (0.75 arbitrary units (AUs) vs. 1.30 AUs; P= 0.008). The median of observation of BC patients enrolled in the study was 54 months (1-325). OS of BC patients with the ERP29 AG + GG combined genotypes was higher than that observed in those with the ERP29 AA wild genotype (67.5% vs. 59.0% at 120 months of follow up; P= 0.04). Conclusions: Our data suggest, for the first time, that ERP29 c.*293A > G polymorphism alters the risk and prognosis of BC possibly due to variation in the protein production. Disclosure: All authors have declared no conflicts of interest.

1119PPOLYMORPHISMS IN THE APOPTOSIS PATHWAY IN CUTANEOUS MELANOMA: RECURRENCE AND SURVIVAL

ABSTRACT Aim: Cutaneous melanoma (CM) is notorious for its poor prognosis and resistance to conventional chemotherapy. Genes involved in the apoptosis pathway, such as P53, (tumor suppressor gene), MDM2 (p53 inhibitor), BAX (proapoptotic) and BCL2 (anti-apoptotic) are important for CM growth and survival. The aim of this study was to evaluate whether P53 Arg72Pro, MDM2 T309G, BAX G(-248)A and BCL2 C(-938)A polymorphisms, involved with inherited variations in apoptosis, are associated with relapse free survival (RFS) and overall survival (OS) of CM patients. Methods: Our analysis included 234 consecutive CM patients at diagnosis seen at our University Hospital from December 1989 to November 2013. Genomic DNA from peripheral blood of patients was analyzed by polymerase chain reaction followed by enzymatic digestion for discrimination of pertinent genotypes. RFS and OS were calculated using the univariate Kaplan-Meier estimate probabilities, and differences between survival curves were analysed by the log-rank test. RFS was defined as the time to beginning of treatment until the date of the first relapse. OS was calculated from date of first diagnosis until the date of death or last follow-up. Results: The median period of observation of patients in study was 50 months (9-192) for RFS and 58 months (range: 9-285) for OS. We observed at 120 months of segment, that the RFS was higher in patients with BCL2 CC + CA plus MDM2 TT + TG than patients with BCL2 AA plus MDM2 GG genotypes (54% versus 37%, P = 0.01). The OS in CM patients with the BAX GA + AA was higher than in those with the GG genotypes (100% versus 80%, P = 0.01). Moreover, the OS was also higher in patients with BAX GA + AA plus BCL2 CC + CA combined genotype than in those with BAX GG plus BCL2 AA (100% versus 78%, P = 0.02) and BAX GA + AA plus MDM2 TT + TG than BAX GG plus MDM2 GG (100% versus 71%, P = 0.001). Conclusions: The data suggest, for the first time, that BAX G(-248)A polymorphism may independent or jointly with BCL2 C(-938)A and MDM2 T309G polymorphisms modulate RFS and OS in CM patients. Additional studies will provide some promising guidance for clinical management and tailored or personalized therapeutics in treating for CM. Disclosure: All authors have declared no conflicts of interest.