RESUMO
<p><b>OBJECTIVE</b>To investigate the effect of ASCT2 gene (glutamine transporter) knock-down by shRNA on biological behaviors of colorectal cancer cells.</p><p><b>METHODS</b>shRNA was transfected into colorectal cancer cells Lovo and SW480 to knockdown ASCT2 mediated by Lipofectamine 2000. Reverse transcription-PCR and Western blot were used to examine the mRNA and protein expression of ASCT2. MTT and transwell assay were used to determine the proliferation and invasiveness of Lovo and SW480 cells. Radioactive-tracer was used to detect the uptake of glutamine.</p><p><b>RESULTS</b>ASCT2 mRNA and protein levels were significantly down-regulated by shRNA in Lovo and SW480 cells(P<0.01). MTT and transwell assays showed that ASCT2 knock-down could significantly inhibit the proliferation of Lovo and SW480 cells (A490) and decrease the number of invasive Lovo and SW480 cells from the membrane (both P<0.01). The number of membrane Lovo cells in shASCT group and control group was 46.3±5.9 and 197.7±9.1, respectively while the number of membrane SW480 cells in shASCT group and control group was 29.7±3.8 and 139.0±9.5, respectively. Radioactive-tracer showed that shASCT2 transfection could significantly reduce the uptake of glutamine, with an inhibition rate of 79.15% in Lovo and 67.22% in SW480 cells (both P<0.01).</p><p><b>CONCLUSIONS</b>ASCT2 plays an oncogenic role in colonic cancer, and its promotion mechanism may be associated with glutamine metabolism. ASCT2 may be a novel therapeutic target of colonic cancer.</p>
Assuntos
Humanos , Sistema ASC de Transporte de Aminoácidos , Genética , Fisiologia , Linhagem Celular Tumoral , Fisiologia , Proliferação de Células , Genética , Neoplasias Colorretais , Genética , Regulação para Baixo , Técnicas de Silenciamento de Genes , Métodos , Glutamina , Genética , Fisiologia , Antígenos de Histocompatibilidade Menor , Genética , Fisiologia , Invasividade Neoplásica , Genética , Oncogenes , Genética , RNA Mensageiro , Fisiologia , RNA Interferente Pequeno , Farmacologia , TransfecçãoRESUMO
Glutamine, the most abundant amino acid in bloodstream, is the preferred fuel source for enterocytes. Glutamine exerts its functions through the activity of its transporters, which are located in cytomembrane, to transport it into or out of intestinal epithelial cells. Intestine is the primary center for glutamine metabolism in the body. As ASCT2 and B(0)AT1 are the most important glutamine transporters in the intestine, it wound be helpful to gain the knowledge of the structure, function, and pathologic changes and control strategy of the two transporters in order to have a better understanding of the metabolism and function of glutamine.
Assuntos
Humanos , Sistema ASC de Transporte de Aminoácidos , Transporte Biológico , Enterócitos , Células Epiteliais , Metabolismo , Patologia , Glutamina , Metabolismo , Intestino Delgado , MetabolismoRESUMO
Atualmente, os problemas encontrados no diagnóstico de células escamosas atípicas de significado indeterminado são devido à falta de uniformização dos critérios citológicos utilizados por diferentes observadores em diferentes laboratórios, gerando altas taxas de discrepância entre seus resultados. O objetivo deste trabalho foi revisar os critérios morfológicos para ASC-US e comparar os resultados encontrados por dois diferentes observadores em 45 esfregaços, previamente diagnosticados por dois farmacêuticos bioquímicos especialistas em citologia clínica. Foram revisados 45 esfregaços com ASC-US. Destes, 35 esfregaços tiveram o diagnóstico final conclusivo para ASC-US, confirmado pelos observadores (Índice de concordância de 77,8 por cento; três esfregaços foram considerados pelos observadores apenas, como processos inflamatórios inespecíficos (6,7 por cento).