ABSTRACT
Introduction: Cytogenetics is the area of genetics that studies chromosomes, including numerical changes, and their relationship to structural imbalances. Among the classical cytogenetics tests, the GTG banding karyotype is the most widely used. The period of culture establishment is a critical step, which can affect the pre-analytical phase of the test. Objective: To evaluate, at different establishment times, culture viability and banding resolution. Material and methods: Collection of 10 ml blood from 10 subjects was carried out for culture analysis. For viability analysis, mitotic index (MI) and banding resolution were assessed. Results: The comparative analysis of MI showed significant difference between times. In the assessment of banding resolution, the mean value of the bands was higher at times zero and 24 hour. Discussion: The MI reflects inhibition of cell cycle progression and/or loss of ability to proliferate. When the pair analysis was performed, a difference between zero and 48 hours was observed. The average number of bands analyzed at times zero and 24 hours did not indicate difference in the quantity and quality of the bands when cultures were grown immediately after blood collection or within 24 hours. At the 48th hour after blood collection significant reduction of band resolution was observed. Conclusion: These data highlight the importance of the biological material quality, as viability is lower when the culture is grown after 24 hours, as well as the banding resolution...
Introdução: Citogenética é a área da genética que estuda os cromossomos, incluindo alterações numéricas, estruturais e sua relação com distúrbios. Nos exames de citogenética clássica, o cariótipo com bandamento GTG é a metodologia mais utilizada, sendo o período de implantação da cultura uma fase crítica, que pode comprometer a fase pré-analítica do exame. Objetivo: Avaliar, em diferentes tempos de implantação, a viabilidade das culturas e o número de bandas. Material e métodos: Foram colhidos 10 ml de sangue de 10 sujeitos para realização das culturas. Para análise da viabilidade, foram utilizados o índice mitótico (IM) e a análise de resolução por bandamento. Resultados: A comparação dos valores do IM demonstrou desigualdade entre os diferentes tempos analisados. Na análise de resolução, o valor médio das bandas foi maior nos tempos zero e 24 horas. Discussão: O IM reflete a inibição da progressão do ciclo celular e/ou a perda da capacidade de proliferação. Quando o IM foi comparado dois a dois, verificou-se a diferença entre os tempos zero e 48 horas. O número médio de bandas analisadas nos tempos de zero e 24 horas mostrou que não houve diferença na quantidade e na qualidade das bandas quando as culturas foram implantadas logo após a coleta do sangue ou em até 24 horas. No tempo de 48 horas, houve significativa redução da resolução. Conclusão: Os dados obtidos reforçam a importância da qualidade do material biológico, visto que a viabilidade decai quando a implantação da cultura é realizada após 24 horas, assim como a resolução das bandas...
Subject(s)
Humans , Male , Female , Adult , Cytogenetic Analysis/methods , Mitotic Index/methods , Karyotyping , Time FactorsABSTRACT
Laryngeal squamous cell carcinoma (LSCC) is a malignant neoplasm exhibiting aggressive phenotype, high recurrence rate, and risk of developing second primary tumors. Current evidence suggests that cells in the invasive front of carcinomas have different molecular profiles compared to those in superficial areas. This study aimed to identify candidate genes in the invasive front and superficial cells from laryngeal carcinomas that would be useful as molecular markers. Invasive front and tumor surface cells of 32 LSCC were evaluated by high-resolution comparative genomic hybridization. Both CCND1 copy number gains and cyclin D1 protein expression were evaluated to confirm gains of 11q13.3. Losses of 3q26.2-q29 and 18q23 were confirmed by loss of heterozygosity analysis. The most frequent chromosomal alterations observed only in invasive front cells involved gains of 1p, 4q, and 9p and losses of 3p, 11p, 12p, 13q, 17q, 18p, 19q, 20q, 21q, and Xp. Gains of 11q13 were detected in both components from glottis and supraglottis but only in invasive front cells from transglottic tumors. Fluorescence in situ hybridization confirmed gains of CCND1/CPE11 in a subset of cases. In supraglottic tumors, cyclin D1 positivity was associated with distant metastasis (P = 0.0018) and with decreased disease-free survival (P = 0.042). Loss of heterozygosity at 3q26.2 and 18q23 were associated with lymph node involvement (P = 0.055) and worsened prognosis, respectively. In conclusion, this study revealed regions that could be targeted in the search for molecular markers in LSCC. Cyclin D1 may be useful as a prognostic marker in supraglottic tumors.
Subject(s)
Carcinoma, Squamous Cell/genetics , Chromosome Aberrations , Laryngeal Neoplasms/genetics , Carcinoma, Squamous Cell/secondary , Chromosomes, Human , Comparative Genomic Hybridization , Cyclin D1/metabolism , Female , Gene Dosage , Humans , Immunohistochemistry , Laryngeal Neoplasms/pathology , Laser Capture Microdissection , Loss of Heterozygosity , Lymphatic Metastasis , Male , Microsatellite Instability , Microsatellite Repeats , Middle Aged , Tissue Array AnalysisABSTRACT
The cortactin gene, mapped at 11q13, has been associated with an aggressive clinical course in many cancers because of its function of invasiveness. This study evaluated CTTN protein and its prognostic value in the deep invasive front and superficial areas of laryngeal squamous cell carcinomas. The transcript expression levels were evaluated in a subset of cases. Overexpression of CTTN cytoplasmatic protein (80% of cases in both the deep invasive front and superficial areas) and transcript (30% of samples) was detected in a significant number of cases. In more than 20% of cases, observation verified membrane immunostaining in the deep invasive front and superficial areas. Perineural invasion was significantly associated with N stage and recurrence (P = .0058 and P = .0037, respectively). Higher protein expression levels were correlated with perineural invasion (P = .004) in deep invasive front cells, suggesting that this area should be considered a prognostic tool in laryngeal carcinomas. Although most cases had moderate to strong CTTN expression on the tumor surface, 2 sets of cases revealed a differential expression pattern in the deep invasive front. A group of cases with absent to weak expression of CTTN in the deep invasive front showed good prognosis parameters, and a second group with moderate to strong expression of CTTN were associated with an unfavorable prognosis, suggesting an association with worse outcome. Taken together, these results suggest that the deep invasive front might be considered a grading system in laryngeal carcinomas and that cortactin is a putative marker of worse outcome in the deep invasive front of laryngeal carcinomas.
Subject(s)
Biomarkers, Tumor/genetics , Carcinoma, Squamous Cell/genetics , Cortactin/genetics , Laryngeal Neoplasms/genetics , Neoplasm Invasiveness/genetics , Carcinoma, Squamous Cell/metabolism , Carcinoma, Squamous Cell/pathology , Female , Gene Amplification , Humans , Laryngeal Neoplasms/pathology , Male , Middle Aged , Neoplasm Invasiveness/pathology , PrognosisABSTRACT
Câncer de laringe ocorre em 25 dos carcinomas de cabeça e pescoço e compreende 2 de todas as doenças malignas. É comum o aparecimento de segundos tumores primários e, aproximadamente, 5 dos pacientes apresentam cânceres sincrônicos. Há várias evidências indicando que a população celular presente no fronte de invasão possui características moleculares diferentes das áreas tumorais superficiais, tornando esta região importante para avaliação prognóstica. Neste estudo foram investigadas por CGH cromossômico de alta resolução (HR-CGH) as alterações genômicas na área superficial e no fronte de invasão de carcinomas de laringe e selecionadas regiões específicas para serem avaliadas por outras metodologias para a confirmação dos resultados. O componente superficial e o fronte de invasão de 33 carcinomas de laringe fixados em formalina e em blocos de parafina foram avaliados por HR-CGH. Foram detectadas alterações comuns aos dois componentes assim como alterações exclusivas a cada um deles. Adicionalmente, foi investigada e confirmada a expressão aumentada da proteína ciclina D1 o gene CCND1 esta mapeada em 11q13) por análise de expressão em plataformas de microarranjos de tecidos contendo as áreas do tumor e do fronte de invasão. Foi realizada também a análise de marcadores polimórficos de microssatélites mapeados em 3q e 18q em um grupo independente de 33 amostras (DNA tumoral e do sangue periférico) cujos resultados confirmaram as perdas encontradas nestas regiões cromossômicas. A expressão do gene CTTN (mapeada em 11q13) e de sua proteína foram avaliadas e revelaram que os altos níveis de expressão proteica foram correlacionados com invasão perineural nas células do fronte de invasão, sugerindo que esta área pode ser considerada como ferramenta prognóstica em carcinomas de laringe. Foram investigados os ganhos detectados em 2q24...
Subject(s)
Humans , Male , Female , Cortactin , Laryngeal Neoplasms/genetics , Neoplasms, Squamous Cell/geneticsABSTRACT
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.
