RESUMEN
Hemangiosarcoma (HSA) is a malignant neoplasm that occurs in humans and canines with a poor prognosis owing to metastatic spread, despite effective treatment. The frequency of spontaneous HSA development is higher in canines than in humans. Therefore, canine HSA is a useful model of intractable human disease, which requires early detection and an effective therapeutic strategy. A high frequency of the p110α phosphatidylinositol4,5bisphosphate 3kinase catalytic subunit alpha (PIK3CA) mutations is detected in a comprehensive genomewide analysis of canine cases of HSA. The present cloned the fulllength cDNA of canine PIK3CA and identified a mutation in codon 1047 from canine cases of HSA and cell lines that were established from these. The enforced expression of the 1047th histidine residue (H1047)R or L mutants of canine PIK3CA in HeLa cells enhanced epidermal growth factor receptor (EGFR) signaling via Akt phosphorylation. PIK3CA mutant canine HSA cell lines exhibited the hyperphosphorylation of Akt upon EGF stimulation as well. Alpelisib, a molecular targeted drug against PIK3CA activating mutations, exerted a significant antitumor effect in canine PIK3CAmutated HSA cell lines. By contrast, it had no significant effect on canine mammary gland tumor cell lines harboring PIK3CA mutations. On the whole, the findings of the present study suggest that alpelisib may be highly effective against PIK3CA mutations that occur frequently in canine HSA.
Asunto(s)
Hemangiosarcoma , Animales , Línea Celular Tumoral , Fosfatidilinositol 3-Quinasa Clase I/genética , Perros , Células HeLa , Hemangiosarcoma/tratamiento farmacológico , Hemangiosarcoma/genética , Hemangiosarcoma/metabolismo , Humanos , Mutación , TiazolesRESUMEN
Most male dogs are castrated at young ages, making them easy to rear following androgen deprivation. Although the incidence of canine prostate cancer is low, several patients have resistance to androgen therapy and poor clinical prognosis. These outcomes are similar to those of end-stage human androgen-independent prostate cancer. The androgen receptor (AR) of canines has two polyglutamine (polyQ) sequences (Q × 10 and Q × 23) at its N-terminal. The length of polyQ may be a risk factor for the development of prostate cancer in dogs; however, there is no evidence to support this. Hence, we artificially created polyQ deletion mutants of canine AR and evaluated their effects on AR signalling. The deletions of Q × 10 and Q × 23 were associated with significant reductions in AR signalling intensities. The Q × 10 mutants, which increase or decrease Q sequentially, also altered AR signalling. Furthermore, the Q × 10 deletion mutant, compared with the Q × 10 control, altered the intensities of the binding of polyQ to the C-terminal of AR, which contains a ligand-binding domain; this was not observed with the Q × 9, 11, and 12 variants. The number of glutamines in the N-terminals of canine ARs may influence AR signalling intensities and contribute to the risk of prostate cancer in dogs.
Asunto(s)
Enfermedades de los Perros , Neoplasias de la Próstata , Antagonistas de Andrógenos , Andrógenos , Animales , Perros , Glutamina , Humanos , Masculino , Neoplasias de la Próstata/veterinaria , Receptores Androgénicos/genéticaRESUMEN
Isocitrate dehydrogenase 1 (IDH1) mutations are common in gliomas, acute myeloid leukemia, and chondrosarcoma. The mutation 'hotspot' is a single arginine residue, R132. The R132H mutant of IDH1 produces the 2-hydroxyglutarate (2-HG) carcinogen from α-ketoglutarate (α-KG). The reduction of α-KG induces the accumulation of hypoxia-inducible factor-1α subunit (HIF-1α) in the cytosol, which is a predisposing factor for carcinogenesis. R132H is the most common IDH1 mutation in humans, but mutations at the R132 residue can also occur in tumor tissues of dogs. The current study reported the discovery of a novel Tyr208Cys (Y208C) mutation in canine IDH1 (cIDH1), which was isolated from 2 of 45 canine chondrosarcoma cases. As the genomic DNA isolated from chondrosarcoma tissue was mutated, but that isolated from blood was not, Y208C mutations were considered to be spontaneous somatic mutations. The isocitrate dehydrogenase activity of the Y208C mutant was attenuated compared with that of wild-type (WT) cIDH1, but the attenuation of Y208C was less intense than that of the R132H mutation. The induction of HIF-1α response element activity and cell retention of HIF-1α were not increased by Y208C overexpression. In silico and cell biological analysis of IDH1 dimerization revealed that the Y208C mutation, but not the R132H mutation, attenuated binding activity with WT cIDH1. These data suggested that the attenuation of dimerization by the Y208C mutation may cause tumorigenesis through different mechanisms other than via 2-HG production by the IDH1 R132 mutation.
RESUMEN
The objective of this study was to evaluate blood levels of various hormones and compounds related to energy metabolism in cows with subacute ruminal acidosis (SARA). We investigated 11 lactating cows presumed to have SARA based on duration of ruminal pH <5.6 and reticulum pH <6.3 in 2015-2016. Kraft pulp (KP) was used to supplement feed of 7 of the cows studied in an effort to reduce SARA. We continuously monitored ruminal pH and measured blood concentrations of hormones and metabolites related to energy metabolism. Blood measurements included glucose (GLU), total cholesterol (TC), free fatty acid (FFA), insulin, adiponectin (ADN), malate dehydrogenase (MDH), and lactate dehydrogenase (LDH). Additionally, we analyzed milk data (milk yield, milk fat percentage, milk protein percentage, milk urea nitrogen, and protein fat ratio) and reproduction data. The results demonstrated that ADN levels at 4 weeks post-parturition correlated with the total amount of time that the ruminal or reticulum fluid pH was under the threshold during 1 week post-parturition, as well as the numbers of days the cows were diagnosed with SARA (SARA-positive days) up to 30 days post-parturition. SARA-positive days in 2016 were higher than those in 2015. In both years, numbers of SARA-positive days for cows supplemented with KP were lower than those for cows without KP. Increased ADN levels may be a compensatory reaction to frequent SARA which modulates the inflammatory response against high LPS levels and improves insulin resistance caused by LPS. ADN may serve as an estimative index for SARA.