RESUMO
Colorectal cancer is among the most lethal of malignancies, due to its propensity to metastatic spread and multifactorial-chemoresistance. The latter property supports the need to identify novel therapeutic approaches for the treatment of colorectal cancer. MicroRNAs are endogenous non-coding small RNA molecules that function as post-transcriptional regulators of gene expression. Recently, programmed cell death 4 has been identified as a protein that increases during apoptosis. This gene is among the potential targets of miR-21 (OncomiR). Locked nucleic acid-modified oligonucleotides have recently emerged as a potential therapeutic option for targeting microRNAs. The aim of this study was to explore the functional role of locked nucleic acid-anti-miR-21 in the LS174T cell line in vitro and in vivo models. LS174T cells were treated with locked nucleic acid-anti-miR-21 for 24, 48, and 72 h in vitro. The expression of miR-21 and PDCD4 at messenger RNA (mRNA) level was evaluated by quantitative real-time polymerase chain reaction, while the protein level of PDCD4 was determined by Western blotting. Cell migratory behavior and the cluster-forming ability of cells were assessed before and after therapy. The disseminated tumor cells were assessed in the chick chorioallantoic membrane model by Alu quantitative polymerase chain reaction. Locked nucleic acid-anti-miR-21 was transfected successfully into the LS174T cells and inhibited the expression of miR-21. Locked nucleic acid-anti-miR-21 inhibited the migration and the number of cells forming clusters. Moreover, we found that locked nucleic acid-anti-miR-21 transfection was associated with a significant reduction in metastatic properties as assessed by the in ovo model. Our findings demonstrated the novel therapeutic potential of locked nucleic acid-anti-miR-21 in colon adenocarcinoma with high miR-21 expression.
Assuntos
Proteínas Reguladoras de Apoptose/genética , Neoplasias Colorretais/genética , Neoplasias Colorretais/terapia , MicroRNAs/antagonistas & inibidores , MicroRNAs/genética , Oligonucleotídeos/genética , Proteínas de Ligação a RNA/genética , Animais , Proteínas Reguladoras de Apoptose/metabolismo , Linhagem Celular Tumoral , Movimento Celular/genética , Embrião de Galinha , Neoplasias Colorretais/metabolismo , Neoplasias Colorretais/patologia , Terapia Genética , Humanos , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/prevenção & controle , Neoplasias Hepáticas/secundário , MicroRNAs/metabolismo , Metástase Neoplásica , Oligonucleotídeos/administração & dosagem , Proteínas de Ligação a RNA/metabolismo , TransfecçãoRESUMO
OBJECTIVE(S): The oral streptococci especially mutans Streptococci are related with the development of caries in humans. However, they are commonly distributed not only in populations with moderate or high caries incidence but also in populations having no or low caries experience. So, in this study, the differences between protein profiles of Streptococcus mutans from different sources were compared. MATERIALS AND METHODS: Twelve S. mutans strains were isolated from caries-susceptible, caries-free subjects and also S. mutansATCC35668. Total proteins of the strains were extracted, and were analyzed through SDS-PAGE technique with coomassie staining. RESULTS: The protein profiles of caries-susceptible subjects were different from those of caries-free subjects. The major protein bands of SDS-PAGE analysis were observed 26-100kDa and 45-57kDa in caries-susceptible and caries-free subjects, respectively. The major protein bands of SDS-PAGE analysis of S.mutans ATCC35668 were between 35-100kDa. Major significant differences between the protein patterns of two subject groups and interestingly, non-significant differences between the strains of each group were also found. The significant differences between the protein band number of S.mutansATCC35668 and caries-free isolates were also observed; while, there were no significant differences between S.mutansATCC35668 and caries-susceptible subject isolates. The analysis of clusters by Hierarchical cluster method confirmed the clear distinction between S.mutans strains of two groups. Also, there was a clear distinction between the strain of S.mutansATCC35668 and caries free strains CONCLUSION: The less protein bands and diversity in caries-free rather than caries-susceptible isolates may be the less cariogenicity it may cause. It is also revealed that the protein pattern analysis of S.mutans strains can be the suitable way for differentiation of them.
RESUMO
This study was evaluated the clonal diversity of Streptococcus mutans in caries-free and caries-active subjects using MLEE. Strains from caries-free subjects were grouped in a single taxon. Unrooted dendrogram showed that different strains clustered in four different clades, also showed that more than one clonal type can be found in a same individual.
Assuntos
Feminino , Humanos , Masculino , Adulto Jovem , Técnicas de Tipagem Bacteriana/métodos , Portador Sadio/microbiologia , Cárie Dentária/microbiologia , Eletroforese/métodos , Enzimas/análise , Infecções Estreptocócicas/microbiologia , Streptococcus mutans/classificação , Análise por Conglomerados , Fenótipo , Streptococcus mutans/enzimologia , Streptococcus mutans/isolamento & purificaçãoRESUMO
This study was evaluated the clonal diversity of Streptococcus mutans in caries-free and caries-active subjects using MLEE. Strains from caries-free subjects were grouped in a single taxon. Unrooted dendrogram showed that different strains clustered in four different clades, also showed that more than one clonal type can be found in a same individual.
