Activation of MEK1 or MEK2 isoform is sufficient to fully transform intestinal epithelial cells and induce the formation of metastatic tumors

<p>Abstract</p> <p>Background</p> <p>The Ras-dependent ERK1/2 MAP kinase signaling pathway plays a central role in cell proliferation control and is frequently activated in human colorectal cancer. Small-molecule inhibitors of MEK1/MEK2 are therefore viewed as attractive drug candidates for the targeted therapy of this malignancy. However, the exact contribution of MEK1 and MEK2 to the pathogenesis of colorectal cancer remains to be established.</p> <p>Methods</p> <p>Wild type and constitutively active forms of MEK1 and MEK2 were ectopically expressed by retroviral gene transfer in the normal intestinal epithelial cell line IEC-6. We studied the impact of MEK1 and MEK2 activation on cellular morphology, cell proliferation, survival, migration, invasiveness, and tumorigenesis in mice. RNA interference was used to test the requirement for MEK1 and MEK2 function in maintaining the proliferation of human colorectal cancer cells.</p> <p>Results</p> <p>We found that expression of activated MEK1 or MEK2 is sufficient to morphologically transform intestinal epithelial cells, dysregulate cell proliferation and induce the formation of high-grade adenocarcinomas after orthotopic transplantation in mice. A large proportion of these intestinal tumors metastasize to the liver and lung. Mechanistically, activation of MEK1 or MEK2 up-regulates the expression of matrix metalloproteinases, promotes invasiveness and protects cells from undergoing anoikis. Importantly, we show that silencing of MEK2 expression completely suppresses the proliferation of human colon carcinoma cell lines, whereas inactivation of MEK1 has a much weaker effect.</p> <p>Conclusion</p> <p>MEK1 and MEK2 isoforms have similar transforming properties and are able to induce the formation of metastatic intestinal tumors in mice. Our results suggest that MEK2 plays a more important role than MEK1 in sustaining the proliferation of human colorectal cancer cells.</p

Resistência a antimicrobianos de bactérias oriundas de ambiente de criação e filés de tilápias do nilo (Oreochromis niloticus) Antibacterial resistance in bacteria from fish pond and Nile tilapia fillets (Oreochromis niloticus)

A resistência de bactérias a antimicrobianos foi determinada em uma piscicultura de tilápias (Oreochromis niloticus) em tanques de terra, sem utilização de antibióticos para profilaxia ou controle de doenças. Foi selecionado um tanque, capturados peixes e coletadas amostras de conteúdo intestinal e superfície dos peixes, água de abastecimento e do tanque, ração, filés de tilápias frescos e congelados. Colônias representativas foram selecionadas e analisadas pelos testes de Gram, catalase, oxidase e oxidaçãofermentação. Foram selecionadas 89 amostras e submetidas a antibiograma, utilizando vários antimicrobianos. A maioria das bactérias pertenceu às famílias Enterobacteriaceae e Vibrionaceae. Tanto no ambiente de criação como nos filés de tilápias observou-se que os isolados bacterianos apresentaramse resistentes principalmente a ampicilina e eritromicina. O índice de múltipla resistência a antimicrobianos (MAR) foi calculado, sendo que do total de 89 isolados analisados 74 (83%), apresentaram MAR ³ 0,2, ou seja apresentaram-se resistentes a dois ou mais antimicrobianos. As freqüências de índice MAR foram altas e maiores na ração.<br>This study was conducted in a freshwater tilapia farm that has not used any antibiotic. It was selected one pond, caught 15 fishes and collected samples of intestinal content and mucus surface, water influent and pond water, ration, fresh tilapia fillets and frozen fillets.. Phenotypical characteristics, Gram stain, oxidase production, oxidative-fermentative utilization of glucose (O-F) were determined of representative colony. Were selected 89 strains and submitted for antimicrobial sensitivy test using several antibiotics. The major identified bacterial families were belonged Enterobacteriaceae and Vibrionaceae. The most isolates showed resistance to ampicilin and eritromicin. From the 89 isolates evaluated 74 (83%) showed a multiple antibiotic resistance index (MAR) ³ 0.2, that mean resistance to two or more antibiotics. The MAR índice frequency were higher and bigger in the rations