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Cecília Fernandes Lorea,
Daniel Antunes Moreno,
Kleiton Silva Borges,
Carlos Eduardo Martinelli,
Sonir Roberto Rauber Antonini,
Margaret de Castro,
Silvio Tucci,
Luciano Neder,
Leandra Naira Zambelli Ramalho,
Izilda Cardinalli,
Ana Luiza Seidinger,
Maria José Mastellaro,
José Andres Yunes, Silvia Regina Brandalise,
Luiz Gonzaga Tone,
Carlos Alberto Scrideli
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ABSTRACT: Impaired apoptosis has been implicated in the development of childhood adrenocortical tumors (ACT), although the expression of apoptosis-related gene expression in such tumors has not been reported.
The mRNA expression levels of the genes CASP3, CASP8, CASP9, FAS, TNF, NFKB, and BCL2 were analyzed by quantitative real-time PCR in consecutive tumor samples obtained at diagnosis from 60 children with a diagnosis of ACT and in 11 non-neoplastic adrenal samples. BCL2 and TNF protein expression was analyzed by immunohistochemistry.
A significant association was observed between tumor size ≥100 g and lower expression levels of the BCL2 (P=0.03) and TNF (P=0.05) genes; between stage IV and lower expression levels of CASP3 (P=0.008), CASP9 (P=0.02), BCL2 (P=0.002), TNF (P=0.05), and NFKB (P=0.03); Weiss score ≥3 and lower expression of TNF (P=0.01); unfavorable event and higher expression values of CASP9 (P=0.01) and lower values of TNF (P=0.02); and death and lower expression of BCL2 (P=0.04). Underexpression of TNF was associated with lower event-free survival in uni- and multivariate analyses (P<0.01). Similar results were observed when patients with Weiss score <3 were excluded.
This study supports the participation of apoptosis-related genes in the biology and prognosis of childhood ACT and suggests the complex role of these genes in the pathogenesis of this tumor.
European Journal of Endocrinology 05/2012; 167(2):199-208. · 3.42 Impact Factor
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ABSTRACT: The present study aimed to analyze the expression profile of the microRNAs previously described as associated with childhood ALL, miR-92a, miR-100, miR-125a-5p, miR-128a, miR-181b, miR-196b and let-7e, and their association with biological/prognostic features in 128 consecutive samples of childhood acute lymphoblastic leukemia (ALL) by quantitative real-time PCR. A significant association was observed between higher expression levels of miR-196b and T-ALL, miR-100 and patients with low white blood cell count at diagnosis and t(12;21) positive ALL. These findings suggest a potential activity of these microRNAs in pediatric ALL biology.
Leukemia research 11/2011; 36(3):293-8. · 2.36 Impact Factor
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ABSTRACT: Oral mucositis is a common collateral effect among the secondary complications resulting from chemotherapy. The objective of this study was to prospectively evaluate the association of HSV-1, Candida spp., and oral bacteria on the severity of oral mucositis in pediatric acute lymphoblastic leukemia (ALL).
Seventy-one prospective patients were included. Analyses of oral microbiota were conducted on days 14 (D14) and 56 (D56) of the Brazilian GBTLI-99 treatment protocol. Herpes simplex virus (HSV) identification was performed by PCR followed by DNA sequencing analysis. Bacteria and fungi identification was obtained by standard microbiological culture tests.
HSV-1 was found in 10.37% of individual patient samples. One sample was positive for HSV-4. On D14, we found an association between the severity of mucositis and the presence of HSV (p = 0.0347) and Candida spp. (p = 0.0078). At D56, we found an association between the severity of mucositis and the presence of HSV on D14 (p < 0.0001) and HSV presence (p = 0.0317).
The presence of HSV, mainly HSV-1, and Candida spp. was associated with mucositis severity in pediatric ALL. No association could be found between bacterial CFU and severity of mucositis.
Supportive Care in Cancer 05/2011; 20(5):1101-7. · 2.09 Impact Factor
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ABSTRACT: The interactions of acute lymphoblastic leukemia (ALL) blasts with bone marrow (BM) stromal cells have a positive impact on leukemia cell survival. In the present study, we proposed to identify and investigate the role of molecules critically involved in leukemia--microenvironment crosstalk.
Gene expression profiling analyses of BM mesenchymal stem cells (BMMSC) were performed following stimulation by ALL cells. CCL2 and IL-8 plasma levels were evaluated from ALL patients and controls. Expression of the CCL2 and IL-8 receptors in ALL was determined by RT-PCR. The biological effects of CCL2, IL-8 or its neutralizing antibodies in primary precursor-B ALL and BMMSC cells were evaluated using in vitro assays.
Leukemia stimulation of BMMSC upregulated the expression of several inflammatory chemokines, including CCL2 and IL-8. The BM plasma levels of CCL2 and IL-8 in children at diagnosis were significantly higher than in healthy controls (P < 0.001). Functional studies revealed that CCL2 and IL-8 enhanced the capacity of BMMSC to support adhesion of ALL cells. CCL2 and IL-8 were also found to enhance BMMSC survival and to increase their proliferation. ALL cells were not directly affected by CCL2 or IL-8.
The leukemic BM microenvironment had increased levels of CCL2 and IL-8. These chemokines are known to have suppressive effects in normal hematopoiesis. Our data indicate that CCL2 and IL-8 have a positive impact on BMMSC survival, proliferation, and adhesiveness to ALL cells. Leukemia-associated CCL2 and IL-8 upregulation may represent one possible mechanism of microenvironment perversion in favor of ALL cells.
Pediatric Blood & Cancer 04/2011; 56(4):568-77. · 1.89 Impact Factor
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Juliana G Assumpção,
Ana Luíza Seidinger,
Maria José Mastellato,
Raul C Ribeiro,
Gerard P Zambetti,
Ramapriya Ganti,
Kumar Srivastava,
Sheila Shurtleff,
Deqing Pei,
Luiz Carlos Zeferino,
Rozany M Dufloth, Silvia Regina Brandalise,
José Andres Yunes
BMC Cancer 01/2011; 11:152. · 3.01 Impact Factor
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ABSTRACT: Immunoglobulin (Ig) and T-cell receptor (TCR) gene rearrangements function as specific markers for minimal residual disease (MRD), which is one of the best predictors of outcome in childhood acute lymphoblastic leukemia (ALL). We recently reported on the prognostic value of MRD during the induction of remission through a simplified PCR method. Here, we report on gene rearrangement frequencies and offer guidelines for the application of the technique.
Two hundred thirty-three children had DNA extracted from bone marrow. Ig and TCR gene rearrangements were amplified using consensus primers and conventional PCR. PCR products were submitted to homo/heteroduplex analysis. A computer program was designed to define combinations of targets for clonal detection using a minimum set of primers and reactions.
At least one clonal marker could be detected in 98% of the patients, and two markers in approximately 80%. The most commonly rearranged genes in precursor B-cell ALL were IgH (75%), TCRD (59%), IgK (55%), and TCRG (54%). The most commonly rearranged genes for T-ALL were TCRG (100%) and TCRD (24%). The sensitivity of primers was limited to the detection of 1 leukemic cell among 100 normal cells.
We propose that eight PCR reactions per ALL subtype would allow for the detection of two markers in most cases. In addition, these reactions are suitable for MRD monitoring, especially when aiming the selection of patients with high MRD levels (≥ 10(-2)) at the end of induction therapy. Such an approach would be very useful in centers with limited financial resources.
Pediatric Blood & Cancer 12/2010; 55(7):1278-86. · 1.89 Impact Factor
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ABSTRACT: The inherited, low-penetrance arginine-to-histidine substitution at codon 337 (R337H) of the tumor protein 53 gene (TP53) is clustered in southeast Brazil (estimated frequency, 0.3%). Although its tumorigenic effect initially appeared to be tissue-specific, recent evidence suggests its association with a broader range of tumors. Therefore, the authors of this report investigated the spectrum of pediatric malignancies associated with the TP53 R337H mutation at a single referral institution in southeast Brazil.
Genomic DNA samples from 493 children with malignancies were screened for the R337H mutation. Available tumor samples from carriers were investigated for loss of heterozygosity (LOH) and nuclear p53 accumulation. Clinical data were obtained from medical records.
Sixty-five of 70 patients (93%) with adrenocortical tumors (ACTs), 9 of 13 patients (69%) with choroid plexus carcinoma (CPC), and 3 of 41 patients (7.3%) with osteosarcoma carried the mutation. The proportion of CPC to choroid plexus papilloma (CPP) was much higher than that reported elsewhere. Osteosarcoma in carriers had a significantly poorer outcome (P = .02). The mutation was not identified in patients who had acute lymphoblastic leukemia (ALL) (n = 187), recurrent ALL (n = 49), acute myeloid leukemia (n = 44), lymphoma (n = 30), non-CPC central nervous system tumors (n = 26), Ewing sarcoma (n = 25), or rhabdomyosarcoma (n = 8). Among the tumors that were available for analysis, LOH with retention of the mutant allele was confirmed in 21 of 21 ACTs, in 2 of 2 CPCs, and in 2 of 3 osteosarcomas that were positive for R337H. CPCs and osteosarcomas that were positive for R337H had marked nuclear accumulation of p53.
The current findings demonstrated compellingly that the TP53 R337H mutation is associated not only with ACT but also with CPC and, to a lesser extent, with osteosarcoma, both of which are core-component tumors of the Li-Fraumeni syndrome.
Cancer 12/2010; 117(10):2228-35. · 4.77 Impact Factor
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ABSTRACT: Intensive chemotherapy regimens can result in severe toxicities, particularly those that involve the digestive systems, leading to morbidity and mortality in this group of patients. Acute enterocolitis can be a frequent complication. The authors performed a retrospective review or patients treated at their institution to ascertain the prognostic value of the clinical symptoms and signs of acute enterocolitis, the corresponding abdominal ultrasonographic findings, and the impact of previous chemotherapy. Amongst 1159 patients with cancer treated at the Centro Infantil Boldrini from 2003 to 2007, 188 (16.2%) patients had 1 or more episode of enterocolitis. An intestinal wall thickness of >or=3 mm on ultrasound was considered diagnostic of enterocolitis. There were 231 episodes of enterocolitis with a death rate of 11.7%. Previous therapy with cytarabine and the presence of abdominal distention affected survival. An intestinal wall thickness of >or=10 mm in the ultrasonographic examination was associated with greater mortality. In multivariate analysis, age, gender, tumor type, degree of neutropenia, intestinal wall thickness, and number of intestinal segments were not statistically significant difference. In children and young adults with cancer and enterocolitis, the clinical findings of 4 or more symptoms and presence of abdominal distention were associated with higher risk of death. Use of cytarabine and an intestinal wall thickness of >or=10 mm were associated with a higher death rate.
Pediatric Hematology and Oncology 09/2010; 27(6):462-70. · 0.89 Impact Factor
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ABSTRACT: Altered expression of histone deacetylases (HDACs) is a common feature in several human malignancies and may represent an interesting target for cancer treatment, including haematological malignancies. We evaluated the mRNA gene expression profile of 12 HDAC genes by quantitative real-time polymerase chain reaction in 94 consecutive childhood acute lymphoblastic leukaemia (ALL) samples and its association with clinical/biological features and survival. ALL samples showed higher expression levels of HDAC2, HDAC3, HDAC8, HDAC6 and HDAC7 when compared to normal bone marrow samples. HDAC1 and HDAC4 showed high expression in T-ALL and HDAC5 was highly expressed in B-lineage ALL. Higher than median expression levels of HDAC3 were associated with a significantly lower 5-year event-free survival (EFS) in the overall group of patients (P = 0·03) and in T-ALL patients (P = 0.01). HDAC7 and HADC9 expression levels higher than median were associated with a lower 5-year EFS in the overall group (P = 0.04 and P = 0.003, respectively) and in B-lineage CD10-positive patients (P = 0.009 and P = 0·005, respectively). Our data suggest that higher expression of HDAC7 and HDAC9 is associated with poor prognosis in childhood ALL and could be promising therapeutic targets for the treatment of refractory childhood ALL.
British Journal of Haematology 09/2010; 150(6):665-73. · 4.94 Impact Factor
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ABSTRACT: SUMMARY: Denys-Drash syndrome (DDS, Online Mendelian Inheritance in Man number 194080) is a rare human developmental disease generally occurring in 46,XY individuals characterized by the combination of disorder of sex development, early onset nephropathy, and Wilms' tumor (WT). DDS is mainly caused by mutations in the WT1 gene. This report describes a novel WT1 gene mutation in a DDS patient. Sequencing the WT1 gene revealed a heterozygous transversion CAT>AAT within exon 8, causing the substitution of an asparagine for a histidine at residue 377. The p.H377N mutation is predicted to diminish the WT1 protein DNA-binding affinity as it might disrupt the normal zinc finger 2 conformation.
Journal of Pediatric Hematology/Oncology 08/2010; 32(6):486-8. · 1.16 Impact Factor
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Carlos Alberto Scrideli,
Maria Angélica Abdala Cortez,
José Andres Yunes,
Rosane Gomes de Paula Queiróz,
Elvis Terci Valera,
Juliana França da Mata,
Silvia Regina Caminada Toledo,
Priscila Pavoni-Ferreira,
Maria Lúcia de Martino Lee,
Antonio Sérgio Petrilli, Silvia Regina Brandalise,
Luiz Gonzaga Tone
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ABSTRACT: This study evaluates the mRNA expression profile of genes TIMP1, TIMP2, MMP2 and MMP9 in diagnostic bone marrow samples from 134 consecutive ALL children by real-time quantitative PCR. A significant association was observed between higher expression levels of MMP9 and low risk group and absence of extramedullary infiltration and higher expression levels of TIMP2 and MMP2 with T-ALL. TIMP1 gene expression values higher than the median were associated with a significantly lower 5-year event free-survival in univariable (P=0.04) and multivariable analysis (P=0.01). Our data address new information in the complex interaction of the migration/adhesion genes and childhood ALL.
Leukemia research 10/2009; 34(1):32-7. · 2.36 Impact Factor
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Leukemia research 08/2009; 33(12):e228-9. · 2.36 Impact Factor
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ABSTRACT: Osteosarcomas (OS) are aggressive tumors of the bone and often have a poor prognosis. The tumors exhibit karyotypes with a high degree of complexity, which has made it difficult to determine whether any recurrent chromosomal aberrations characterize OS. To address inherent difficulties associated with classical cytogenetic analysis, comparative genomic hybridization (CGH) was applied to OS tissue. Forty-one pediatric OS specimens were analyzed by a CGH technique: 24 female and 17 male patients, with a median age of 12 years and 4 months. Chromosomal abnormalities were highly diverse and variable, including gains of chromosome 1p, 2p, 3q, 5q, 5p, and 6p and losses of 14q (50% in 14q11.2), 15q, and 16p. A high level of losses of chromosome 21 was present (26/41 cases; P = 0.008), most often loss of the 21q11.2 approximately 21 region. These novel findings in chromosome 21 of pediatric OS tumors suggest that specific sequences mapping to these chromosomal regions are likely to play a role in the development of OS.
Cancer Genetics and Cytogenetics 06/2007; 175(1):35-40. · 1.39 Impact Factor
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ABSTRACT: Approximately 35% of T-cell acute lymphoblastic leukemia (T-ALL) cases have chromosomal translocations as evaluated by conventional cytogenetic methods (G-banding). Some chromosomal translocations are associated with morphologically and immunophenotypically distinct leukemia subtypes and define patients with different clinical outcomes. Chromosomal translocations may deregulate gene expression, thus contributing to the development of neoplasia, either by placing a putative oncogene under the control of strong regulatory elements or by generating chimeric genes and oncogenic fusion proteins. We report here a novel der(12)t(7;12)(p15;q24.3) in a child with T-ALL. Cloning and characterization of the breakpoint region may contribute to the discovery of new genes that are important in T-ALL.
Cancer Genetics and Cytogenetics 06/2004; 151(1):78-81. · 1.39 Impact Factor
