Microallelotyping defines novel regions of loss of heterozygosity in uterine leiomyomas

Uterine leiomyomas are extremely common, benign, smooth muscle tumors that represent a significant public health problem. Although there have been few molecular studies of uterine leiomyomas, most of them have reported a very low frequency of loss of heterozygosity (LOH) in different regions of the genome. The detection of LOH has been used to identify genomic regions that harbor tumor suppressor genes and to characterize different tumor types. We have used a set of 15 microsatellite polymorphism markers to examine the frequency of allele loss in a panel of 64 human uterine leiomyomas matched to normal DNAs. The markers were chosen from regions involved in losses identified by comparative genomic hybridization in a subset of uterine leiomyomas described in a previous report. DNA from tumors and normal tissue was amplified by the polymerase chain reaction and subsequently analyzed using an ABI Prism 377 DNA automated sequencer. The frequency of LOH observed was low, except for the markers D15S87 (15q26.3), D7S493 (7p15.3), and D7S517 (7p22.2). No changes in microsatellite size were detected in our samples. These results provide useful clues for identifying putative tumor suppressor genes associated with a subset of uterine leiomyomas. (C) 2004 Wiley-Liss, Inc

Human papillomavirus detection using PCR and ATR-FTIR for cervical cancer screening

Made available in DSpace on 2019-09-12T16:53:41Z (GMT). No. of bitstreams: 0 Previous issue date: 2018Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)The human papillomavirus (HPV) genital infection is considered one of the most common sexually transmitted diseases worldwide, and has been associated with cervical cancer. The objective of this study was to investigate the efficacy of the diagnostic methods: polymerase chain reaction (PCR) and Fourier transform infrared (FTIR) equipped with an ATR (Attenuated Total Reflectance) unit (Pike Tech) spectroscopy, to diagnose HPV infection in women undergoing gynecological examination. Seventeen patients (41.46%) of the 41 patients analyzed were diagnosed with exophytic/condyloma acuminate lesions by clinical analysis, 29 patients (70.7%) (G1 group) of the 41 patients, showed positive result for HPV cell injury by oncotic colpocitology and 12 patients (293%) (G2 group), presented negative result for cellular lesion and absence of clinical HPV lesion. Four samples were obtained per patient, which were submitted oncotic colpocitology analysis (Papanicolau staining, two samples), PCR (one sample) and ATR-FTIR analysis (one sample). L1 gene was amplified by PCR technique with specific GP5+/GP6+ and MY09/MY11 primers. PCR results were uniformly positive for presence of HPV in all analyzed samples. Multivariate analysis of ATR-FTIR spectra suggests no significant biochemical changes between groups and no clustering formed, concurring with results of PCR. This study suggests that PCR and ATR-FTIR are highly sensitive technique for HPV detection. (C) 2018 Elsevier B.V. All rights reserved.[Rymsza, Taciana; Ribeiro, Eliane Aline; Canevari, Renata de Azevedo] Univ Vale Paraiba, Lab Biol Mol Canc, UNIVAP, Inst Pesquisa & Desenvolvimento, Ave Shishima Hifumi 2911, BR-12244000 Sao Paulo, SP, Brazil[das Chagas e Silva de Carvalho, Luis Felipe] Univ Vale Paraiba, Lab Espect Vibrac Biomed, UNIVAP, Inst Pesquisa & Desenvolvimento, Ave Shishima Hifumi 2911, BR-12244000 Sao Paulo, SP, Brazil[Bhattacharjee, Tanmoy] Univ Vale Paraiba, Lab Nanosensores, UNIVAP, Inst Pesquisa & Desenvolvimento, Ave Shishima Hifumi 2911, BR-12244000 Sao Paulo, SP, Brazil[das Chagas e Silva de Carvalho, Luis Felipe] Universidade de Taubaté (Unitau), Dept Odontol, , Rua Operarios 53, BR-12020270 Sao Paulo, SP, Brazi

ANÁLISE DE EXPRESSÃO GÊNICA NA IDENTIFICAÇÃO DE POTENCIAIS MARCADORES PREDITIVOS EM NEOPLASIAS MALIGNAS MAMÁRIAS

O status dos linfonodos axilares é o fator prognóstico mais informativo no tratamento das pacientes com câncer de mama. Contudo, atualmente, na prática clínica, a decisão da dissecação dos linfonodos é ainda realizada por meio da biopsia do linfonodo sentinela, o que, muitas vezes, pode trazer sequelas para a paciente ou originar resultados incorretos. Assim, a identificação de marcadores moleculares, na neoplasia maligna primária, que possa permitir uma classificação mais precisa das pacientes em relação à necessidade, ou não, da dissecação dos linfonodos axilares é extremamente importante para uma conduta clínica mais adequada. O objetivo deste estudo foi determinar se os genes SERPINA1, TFF3, TFF1, ARD1A, NGX6 e DKK1 são marcadores preditivos em câncer de mama, pela análise de expressão gênica de RT-qPCR. Para isso, foi comparado o grupo de neoplasias malignas primárias com envolvimento de linfonodos com neoplasias malignas primárias sem linfonodos acometidos, além da análise dos linfonodos correspondentes. Para todos os genes avaliados, apenas o gene TFF1 apresentou expressão diferencial na comparação da neoplasia maligna primária com o linfonodo correspondente, embora não apresentou diferença estatística na comparação da neoplasia maligna primária linfonodo positivo com a neoplasia maligna primária linfonodo negativo. Os resultados obtidos demonstraram que nenhum dos genes avaliados pode ser considerado marcador preditivo em câncer de mama

Polymorphisms of CYP17A1, CYP19, and androgen in Brazilian women with uterine leiomyomas

Background: Uterine leiomyomas are common, benign, smooth muscle tumors representing a significant public health problem. The aim of this study was to investigate CYP17A1, CYP19, and androgen (AR) polymorphisms, their relative risks for uterine leiomyomas and possible associations with clinical parameters.Methods: Uterine leiomyoma tissues and blood samples were obtained from 87 patients, as were peripheral blood samples from 68 control women. Clinical data were recorded in both groups. The CYP17A1 (rs743572) polymorphism was analyzed by PCR-RFLP, and the CYP19 [TTTA](n) repeat and AR [CAG](n) repeat were analyzed using PCR-based GeneScan analysis. AR loss of heterozygosity (LOH) and microsatellite instability were also evaluated, while samples exhibiting LOH were analyzed for X inactivation.Results: Clinical parameters related to disease development did not differ between cases and controls. CYP17A1 *A2/*A2 genotype was prevalent in non-white women. CYP17A1, CYP19, and AR genotypes and alleles did not differ between groups. However, alleles presenting [TTTA](7) repeats in intron 4 of CYP19 were more frequent in the control group (p=0.0550). Shorter and longer [CAG]n repeat alleles of AR were exclusive to the leiomyoma group. The LOH assay showed allele losses at AR locus in four informative tumors and X chromosome inactivation analysis revealed that these tumors retained the active allele.Conclusions: The overall lack of association between uterine leiomyomas with polymorphisms involved in steroidogenesis or steroid metabolism is consistent with the hypothesis that these polymorphisms do not substantially contribute to the development of these tumors

Shorter CAG repeat in the AR gene is associated with atypical hyperplasia and breast carcinoma

Background: Previous reports into the role of [CAG]n repeat lengths in the androgen receptor (AR) gene indicate that these may play an important part in the development and progression of breast cancer, however, knowledge regarding benign breast lesions is limited. Patients and Methods: PCR-based GeneScan analysis was used to investigate the [CAG]n repeat length at exon 1 of the AR gene in 59 benign breast lesions (27 fibroadenomas, 18 atypical hyperplasias, and 14 hyperplasias without atypia) and 54 ductal breast carcinomas. Seventy-two cancer-free women were used as a control group. In addition, [CAG]n repeats were evaluated for the presence of loss of heterozygosity (LOH) and microsatellite instability (MSI) in a subset of these samples (27 fibroadenomas, 14 hyperplasias without atypia and 22 breast carcinomas). Results: Shorter [CAG]n repeat lengths were strongly correlated with atypical hyperplasias (p=0.0209) and carcinomas (p<0.0001). LOH was found in 1/12 and 4/20 informative cases of hyperplasias without atypia and breast carcinomas, respectively. Three patients with breast carcinoma who had previously presented atypical hyperplasia showed a reduction in the [CAG]n repeat length in their carcinomas. Conclusion: Short [CAG]n repeat length (≤20) polymorphisms are strongly associated with breast carcinomas and atypical hyperplasias. Although non-significant, a subgroup of patients with breast carcinoma and genotype SS showed an association with parameters of worse outcome

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 observer's 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 CaF2 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)&#174;. 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