5 research outputs found
Evaluation of valosin containing protein (P97) as a cancer biomaker in canine lymphomas
Le lymphome est l'une des tumeurs les plus communes tant chez le chien que lāhumain. Chaque anneĢe, un nombre important de chiens deĢveloppe ce cancer agressif. La majoriteĢ deĢceĢdant un an suivant le diagnostic. Le lymphome canin est maintenant identifieĢ comme un excellent modeĢle de recherche pour la tumeur chez l'homme, particulieĢrement en ce qui concerne la biologie moleĢculaire de la maladie. En conseĢquence, la recherche sur le lymphome canin sera beĢneĢfique non seulement pour les chiens mais aussi pour lāoncologie humaine. Parmi les meĢthodes diagnostiques de choix pour deĢpister de facĢ§on haĢtive le lymphome se trouve la mesure de marqueurs tumoraux. Ceci a lāavantage dāeĢtre peu invasive, simple et peu dispendieuse. Ainsi, dans le but dāeĢvaluer la proteĢine VCP (valosin containing protein) comme biomarqueur tumoral dans les lymphomes canins aĢ cellules B et T, nous avons eĢvalueĢ la proteĢine VCP par immunobuvardage sur seĢrums et tissus tumoraux de chiens atteints et par immunohistochimie sur des tumeurs de haut grade, grade intermeĢdiaire et bas grade. Pour mieux deĢfinir lāexpression de VCP dans les cellules canceĢreuses, nous avons eĢgalement examineĢ par immunobuvardage les niveaux de VCP dans 3 ligneĢes cellulaires: CLBL-1, CL-1, et 17-71. Il sāaveĢre que les lymphomes aĢ cellules B de haut grade avaient une eĢleĢvation significative du taux de VCP compareĢ aux tumeurs de bas grade (P < 0,05). De meĢme, une accumulation importante de VCP a eĢgalement eĢteĢ deĢtecteĢe dans les ligneĢes tumorales compareĢes aux cellules mononucleĢaires du sang peĢripheĢrique (P < 0,05). Dāautre part, le taux seĢrique de VCP est resteĢ similaire aĢ ceux des chiens normaux. Ces reĢsultats suggeĢrent une correĢlation entre le taux de VCP et le degreĢ de maligniteĢ des lymphomes aĢ cellules B. En conclusion, la proteĢine VCP doit faire lāobjet dāune eĢvaluation approfondie pour deĢterminer son utiliteĢ comme marqueur pronostique.Lymphoma is one of the common malignancies in both dogs and humans. Annually, an important number of canine patients develop this aggressive cancer and a majority succumbs to the disease within one year. In recent years, canine lymphoma has been increasingly recognized as an excellent model for the disease in humans, especially with regards to the molecular biology of the disease. Consequently, research targeted at canine lymphoma benefits not only dogs but the field of human oncology as well. Among the most desirable diagnostic and screening tests for lymphoma is the measurement of cancer biomarkers. They have the advantage of being minimally invasive, simple, and inexpensive. Thus, with the aim of evaluating valosin containing protein (VCP) as a cancer biomarker in canine B and T-cell lymphomas, we first performed western blots on sera and tumor tissue of dogs with lymphoma and then immunohistochemical analysis on low, intermediate and high-grade tumors. To further determine VCP expression in cancer cells, we also examined VCP levels by immunoblotting in 3 tumor cell lines: CLBL-1, CL-1, and 17-71. High-grade B-cell lymphomas had significantly increased levels of VCP compared to low-grade tumors (P < 0.05). Additionally, we detected a corresponding accumulation of VCP in tumor cells lines compared to peripheral blood mononuclear cells (PBMCs) (P < 0.05). In contrast, VCP levels were not elevated in sera of dogs with lymphoma compared to healthy controls. These results suggest that VCP positively correlates with malignancy in canine B-cell lymphomas. We conclude that VCP merits further investigation to determine its potential as a clinically useful prognosis biomarker for canine B-cell lymphoma
Significance of the Wnt signaling pathway in coronary artery atherosclerosis
IntroductionThe progression of coronary atherosclerosis is an active and regulated process. The Wnt signaling pathway is thought to play an active role in the pathogenesis of several cardiovascular diseases; however, a better understanding of this system in atherosclerosis is yet to be unraveled.MethodsIn this study, real-time quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blotting were used to quantify the expression of Wnt3a, Wnt5a, and Wnt5b in the human coronary plaque, and immunohistochemistry was used to identify sites of local expression. To determine the pathologic significance of increased Wnt, human vascular smooth muscle cells (vSMCs) were treated with Wnt3a, Wnt5a, and Wnt5b recombinant proteins and assessed for changes in cell differentiation and function.ResultsRT-PCR and Western blotting showed a significant increase in the expression of Wnt3a, Wnt5a, Wnt5b, and their receptors in diseased coronary arteries compared with that in non-diseased coronary arteries. Immunohistochemistry revealed an abundant expression of Wnt3a and Wnt5b in diseased coronary arteries, which contrasted with little or no signals in normal coronary arteries. Immunostaining of Wnt3a and Wnt5b was found largely in inflammatory cells and myointimal cells. The treatment of vSMCs with Wnt3a, Wnt5a, and Wnt5b resulted in increased vSMC differentiation, migration, calcification, oxidative stress, and impaired cholesterol handling.ConclusionsThis study demonstrates the upregulation of three important members of canonical and non-canonical Wnt signaling pathways and their receptors in coronary atherosclerosis and shows an important role for these molecules in plaque development through increased cellular remodeling and impaired cholesterol handling