RESUMO
MicroRNAs (miRNAs) are short non-coding RNA molecules acting as important posttranscriptional gene and protein expression regulators in cancer. The study goal was to examine VEGFA (vascular endothelial growth factor A) expression in hepatocellular carcinoma (HCC) cell lines upon transfection miR-612, miR-637, or miR-874. Methods: MiR-612 mimics, miR-637 mimics, or miR-874 inhibitors were transfected using Lipofectamine RNAiMax in both HCC cell lines, HepG2 and HuH-7. Real-time PCR, Western blotting, and ELISA methods were used to evaluate VEGFA regulation by the miRNAs. Results: Gene and protein expression levels of VEGFA were down-expressed in both cell lines, HepG2 and HuH-7, transfected with miR-612 or miR-637. Transfection with miR-874 inhibitor showed an increase in VEGFA gene expression in HepG2 and HuH-7 cell lines; however, no regulation was observed on VEGFA protein expression by miR-874 inhibition. Correlation analysis between miRNAs and VEGFA protein expression showed that miR-637 and miR-874 expression present inversely correlated to VEGFA protein expression. Conclusions: VEGFA was down-regulated in response to hsa-miR-612 or hsa-miR-637 overexpression; however, the modulation of VEGFA by miR-874 was observed only at the gene expression and thus, needs further investigation.
Assuntos
Carcinoma Hepatocelular , Neoplasias Hepáticas , MicroRNAs , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/patologia , Linhagem Celular , Regulação Neoplásica da Expressão Gênica , Humanos , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/patologia , MicroRNAs/genética , MicroRNAs/metabolismo , Fator A de Crescimento do Endotélio Vascular/genética , Fator A de Crescimento do Endotélio Vascular/metabolismoRESUMO
INTRODUCTION: Liver cirrhosis (LC) is a heterogeneous liver disease, the last stage of liver fibrosis, and the major risk factor for hepatocellular carcinoma (HCC). Our study aimed to evaluate the expression of microRNAs and the endothelial vascular growth factor (VEGFA) gene in LC and HCC. MATERIAL AND METHODS: The sample group consisted of 46 tissue samples: 21 of LC, 15 of HCC, and 10 of non-tumoural and non-cirrhotic liver tissue (control group). MiRNAs were chosen based on a mirDIP prediction database as regulators of the VEGFA gene. Gene expression of VEGF and miRNAs was quantified by real-time quantitative polymerase chain reaction. VEGFA protein expression was evaluated by ELISA. RESULTS: VEGFA gene expression was significantly overexpressed in LC compared to the control group (p < 0.0001). Hsa-miR-206 (p = 0.0313) and hsa-miR-637 (p = 0.0156) were down-expressed in LC. In HCC, hsa-miR-15b (p = 0.0010), hsa-miR-125b (p = 0.0010), hsa-miR-423-3p (p = 0.0010), hsa-miR-424 (p = 0.0313), hsa-miR-494 (p < 0.0001), hsa-miR-497 (p < 0.0001), hsa-miR-612 (p = 0.0078), hsa-miR-637 (p < 0.0001), and hsa-miR-1255b (p = 0.0156) presented down-expression. CONCLUSIONS: Overexpression of VEGFA in LC suggests impairment of angiogenesis in this tissue. The differential expression of microRNAs in LC and HCC observed in our study can lead to the evaluation of possible biomarkers for these diseases.
RESUMO
Os abortamentos espontâneos ocorrem por diversas causas, sendo as anomalias cromossômicas do concepto,as mais freqüentes. Assim, o estudo citogenético de seus produtos e genitores, seguido do aconselhamentogenético aos casais participantes, são condutas essenciais. O presente estudo retrospectivo teve comoobjetivo investigar os cariótipos de 574 amostras de produtos de abortamentos espontâneos, bem comosangue periférico de casais com abortamentos espontâneos recorrentes, para estimar a frequência de alteraçõescromossômicas. Os cariótipos foram previamente realizados a partir da cultura de vilosidade coriônica (abortos)e sangue periférico (casais), seguida da técnica de bandeamento G. Concluiu-se que as frequências observadasde alterações cromossômicas entre os abortos (19,69%) e os casais (7,6%) foram concordantes com a literatura,reforçando a importância da análise citogenética nesses casos.
Miscarriages result of several causes, but chromosomal anomalies are the most frequent. Thus, the cytogeneticstudies of its products and of the parents, followed by genetic counseling to involved couples, are crucialconducts. The present retrospective work aimed to investigate the karyotypes of 574 samples of miscarriageproducts, and the peripheral blood of couples involved with recurrent miscarriages in an attempt to estimatethe frequencies of chromosomal alterations. Karyotypes were previously made from the culture of chorionicvilli (miscarriages) and peripheral blood (couples), followed by the technique of G banding. It was concludedthat the observed frequency of abnormal karyotypes among miscarriage products (19.69%) and couples(7.6%) were both consistent with the literature, reinforcing the importance of cytogenetic study in thesecases.
Assuntos
Aberrações Cromossômicas/estatística & dados numéricos , Aborto Habitual/genética , Análise CitogenéticaRESUMO
As anomalias cromossômicas fazem parte de uma das maiores categorias de doenças genéticas, sendoresponsáveis por diversas malformações congênitas e parte dos insucessos reprodutivos. Este estudo retrospectivo teve como principal objetivo analisar a freqüência de anomalias cromossômicas em 1171indivíduos, encaminhados para exame citogenético, no período de janeiro de 1998 a dezembro de 2007, aoAmbulatório e Laboratório de Genética da Unesp Bauru. Os cariótipos foram previamente realizados com atécnica de bandeamento G em células obtidas a partir de cultura de linfócitos de sangue periférico. Os resultados mostraram 142 casos de cariótipos anormais, sendo as aneuploidias (45,1%) as mais freqüentes,seguidas pelas alterações estruturais (38,7%) e mosaicismos (16,2%). Concluiu-se que esses resultadosconcordam com a literatura pertinente, sendo de alta relevância o estudo citogenético tanto para diagnósticoem indivíduos malformados ou inférteis, como para esclarecer a etiologia da perda fetal e, assim, alertar à possibilidade de futuras malformações congênitas.
Chromosomal abnormalities are part of a larger category of genetic diseases, and responsible for various congenital malformations and part of the reproductive failures. The mainly objective of this retrospective work was to analyze the frequency of chromosomal abnormalities among 1,171 individuals referred to the Ambulatório e Laboratório de Genética, Unesp-Bauru for cytogenetic examination, from January, 1998 to December, 2007. The G-banding technique was previously used to prepare karyotypes of cells from lymphocyte culture of peripheral blood. The results showed 142 cases of abnormal karyotypes. The most frequent were aneuploidies (45.1%), followed by structural anomalies (38.7%), and mosaicisms (16.2%). It was concluded that these results are consistent with those in the literature, and they are important both for the cytogenetic diagnosis in malformed or infertile individuals, and also to clarify the etiology of fetal loss, and thus alert to the possibility of further congenital malformations.