TGF-β1 modulates the homeostasis between MMPs and MMP inhibitors through p38 MAPK and ERK1/2 in highly invasive breast cancer cells

<p>Abstract</p> <p>Background</p> <p>Metastasis is the main factor responsible for death in breast cancer patients. Matrix metalloproteinases (MMPs) and their inhibitors, known as tissue inhibitors of MMPs (TIMPs), and the membrane-associated MMP inhibitor (RECK), are essential for the metastatic process. We have previously shown a positive correlation between MMPs and their inhibitors expression during breast cancer progression; however, the molecular mechanisms underlying this coordinate regulation remain unknown. In this report, we investigated whether TGF-β1 could be a common regulator for MMPs, TIMPs and RECK in human breast cancer cell models.</p> <p>Methods</p> <p>The mRNA expression levels of TGF-β isoforms and their receptors were analyzed by qRT-PCR in a panel of five human breast cancer cell lines displaying different degrees of invasiveness and metastatic potential. The highly invasive MDA-MB-231 cell line was treated with different concentrations of recombinant TGF-β1 and also with pharmacological inhibitors of p38 MAPK and ERK1/2. The migratory and invasive potential of these treated cells were examined in vitro by transwell assays.</p> <p>Results</p> <p>In general, TGF-β2, TβRI and TβRII are over-expressed in more aggressive cells, except for TβRI, which was also highly expressed in ZR-75-1 cells. In addition, TGF-β1-treated MDA-MB-231 cells presented significantly increased mRNA expression of MMP-2, MMP-9, MMP-14, TIMP-2 and RECK. TGF-β1 also increased TIMP-2, MMP-2 and MMP-9 protein levels but downregulated RECK expression. Furthermore, we analyzed the involvement of p38 MAPK and ERK1/2, representing two well established Smad-independent pathways, in the proposed mechanism. Inhibition of p38MAPK blocked TGF-β1-increased mRNA expression of all MMPs and MMP inhibitors analyzed, and prevented TGF-β1 upregulation of TIMP-2 and MMP-2 proteins. Moreover, ERK1/2 inhibition increased RECK and prevented the TGF-β1 induction of pro-MMP-9 and TIMP-2 proteins. TGF-β1-enhanced migration and invasion capacities were blocked by p38MAPK, ERK1/2 and MMP inhibitors.</p> <p>Conclusion</p> <p>Altogether, our results support that TGF-β1 modulates the mRNA and protein levels of MMPs (MMP-2 and MMP-9) as much as their inhibitors (TIMP-2 and RECK). Therefore, this cytokine plays a crucial role in breast cancer progression by modulating key elements of ECM homeostasis control. Thus, although the complexity of this signaling network, TGF-β1 still remains a promising target for breast cancer treatment.</p

Unveiling novel genes upregulated by both rhBMP2 and rhBMP7 during early osteoblastic transdifferentiation of C2C12 cells

<p>Abstract</p> <p>Findings</p> <p>We set out to analyse the gene expression profile of pre-osteoblastic C2C12 cells during osteodifferentiation induced by both rhBMP2 and rhBMP7 using DNA microarrays. Induced and repressed genes were intercepted, resulting in 1,318 induced genes and 704 repressed genes by both rhBMP2 and rhBMP7. We selected and validated, by RT-qPCR, 24 genes which were upregulated by rhBMP2 and rhBMP7; of these, 13 are related to transcription (<it>Runx2, Dlx1, Dlx2, Dlx5, Id1, Id2, Id3, Fkhr1, Osx, Hoxc8, Glis1, Glis3 </it>and <it>Cfdp1</it>), four are associated with cell signalling pathways (<it>Lrp6, Dvl1, Ecsit </it>and <it>PKCδ</it>) and seven are associated with the extracellular matrix (<it>Ltbp2, Grn, Postn, Plod1, BMP1, Htra1 </it>and <it>IGFBP-rP10</it>). The novel identified genes include: <it>Hoxc8, Glis1, Glis3, Ecsit, PKCδ, LrP6, Dvl1, Grn, BMP1, Ltbp2, Plod1, Htra1 </it>and <it>IGFBP-rP10</it>.</p> <p>Background</p> <p>BMPs (bone morphogenetic proteins) are members of the TGFβ (transforming growth factor-β) super-family of proteins, which regulate growth and differentiation of different cell types in various tissues, and play a critical role in the differentiation of mesenchymal cells into osteoblasts. In particular, rhBMP2 and rhBMP7 promote osteoinduction <it>in vitro </it>and <it>in vivo</it>, and both proteins are therapeutically applied in orthopaedics and dentistry.</p> <p>Conclusion</p> <p>Using DNA microarrays and RT-qPCR, we identified both previously known and novel genes which are upregulated by rhBMP2 and rhBMP7 during the onset of osteoblastic transdifferentiation of pre-myoblastic C2C12 cells. Subsequent studies of these genes in C2C12 and mesenchymal or pre-osteoblastic cells should reveal more details about their role during this type of cellular differentiation induced by BMP2 or BMP7. These studies are relevant to better understanding the molecular mechanisms underlying osteoblastic differentiation and bone repair.</p

Estudo da associação entre fatores socioambientais e prevalência de parasitose intestinal em área periférica da cidade de Manaus (AM, Brasil) Study of the association between socio-environmental factors and the prevalence of intestinal parasitosis in the suburbs of the city of Manaus in the state of Amazonas, Brazil

Este estudo teve como objetivo avaliar a associação entre fatores socioambientais e condições de saneamento urbano com a prevalência de parasitoses intestinais, em uma comunidade na periferia da cidade de Manaus. O estudo consistiu de um levantamento socioambiental e de um inquérito parasitológico. Foi encontrada uma comunidade heterogênea, apresentando diferenças em relação aos níveis socioeconômicos e ambientais entre as micro-áreas avaliadas, embora as condições de saneamento básico da comunidade tenham-se mostrado precárias de uma forma geral. A prevalência de parasitose intestinal foi de 44,2%. Não foi identificada diferença significativa entre as micro-áreas com respeito à ocorrência de parasitoses intestinais. Houve associação entre parasitoses intestinais e tipo de construção residencial, faixa etária e procedência da água de higiene pessoal e do lar. O despejo do esgoto a céu aberto foi fator de risco associado a parasitoses intestinais (OR=6,72; p=0,034) e a protozoários intestinais (OR=21,87; p=0,004). Foram fatores de risco, apenas para a presença de protozoários, o despejo de esgoto em igarapé (OR=12,98; p=0,011) e o uso de fossa rudimentar (OR=9,54; p=0,019).<br>This study assesses the association between socio-environmental factors and urban sanitation conditions with the prevalence of intestinal parasitosis in a community on the periphery of the city of Manaus. The study comprised a socio-environmental survey and a parasitological inquiry. A heterogeneous community was revealed with some socio-economic and environmental differences between the micro-areas evaluated, even though the urban sanitation conditions were found to be predominantly precarious. The prevalance of intestinal parasitosis was 44.2%. There was no significant difference between the micro-areas that could explain the occurrence of intestinal parasitosis. An association was found between intestinal parasitosis and residential building types, age bracket and the quality of the water used for personal hygiene and consumption in the home. Open air sewerage was a risk factor associated with intestinal parasitosis (OR=6.72; p=0.034) and also with intestinal protozoa (OR=21.87; p=0.004). In terms of the presence of protozoa, two risk factors were verified: the dumping of sewage directly into the river system (OR=12.98; p=0.011) and the use of rudimentary cesspits (OR=9.54; p=0.019)

Implementation of a multifaceted sepsis education program in an emerging country setting: clinical outcomes and cost-effectiveness in a long-term follow-up study

Purpose: To evaluate whether a multifaceted, centrally coordinated quality improvement program in a network of hospitals can increase compliance with the resuscitation bundle and improve clinical and economic outcomes in an emerging country setting. Methods: This was a pre- and post-intervention study in ten private hospitals (1,650 beds) in Brazil (from May 2010 to January 2012), enrolling 2,120 patients with severe sepsis or septic shock. the program used a multifaceted approach: screening strategies, multidisciplinary educational sessions, case management, and continuous performance assessment. the network administration and an external consultant provided performance feedback and benchmarking within the network. the primary outcome was compliance with the resuscitation bundle. the secondary outcomes were hospital mortality, hospital and ICU length of stay, quality-adjusted life year (QALY) gain, and cost-effectiveness. Results: the proportion of patients who received all the required items for the resuscitation bundle improved from 13 % [95 % confidence interval (CI) 8-18 %] at baseline to 62 % (95 % CI 54-69 %) in the last trimester (p < 0.001). Hospital mortality decreased from 55 % (95 % CI 48-62 %) to 26 % (95 % CI 19-32 %, p < 0.001). Full compliance with the resuscitation bundle was associated with lower risk of hospital mortality (propensity weighted corrected risk ratio 0.74; 95 % CI 0.56-0.94, p = 0.02). There was a reduction in the total cost per patient from 29.3 (95 % CI 23.9-35.4) to 17.5 (95 % CI 14.3-21.1) thousand US dollars from baseline to the last 3 months (mean difference -11,815; 95 % CI -18,604 to -5,338). the mean QALY increased from 2.63 (95 % CI 2.15-3.14) to 4.06 (95 % CI 3.58-4.57). for each QALY, the full compliance saves US$5,383. Conclusions: A multifaceted approach to severe sepsis and septic shock patients in an emerging country setting led to high compliance with the resuscitation bundle. the intervention was cost-effective and associated with a reduction in mortality.Hosp Paulistano, Unidade Terapia Intens, BR-01321001 São Paulo, BrazilUniv São Paulo, Unidade Terapia Intens, Hosp Clin, Disciplina Emergencias Clin, BR-05403000 São Paulo, BrazilLatin Amer Sepsis Inst, BR-04039002 São Paulo, BrazilUniversidade Federal de São Paulo, Escola Paulista Med, Disciplina Anestesiol, BR-04024900 São Paulo, BrazilUniversidade Federal de São Paulo, Escola Paulista Med, Disciplina Anestesiol, BR-04024900 São Paulo, BrazilWeb of Scienc