Transcriptomic profile reveals molecular events associated to focal adhesion and invasion in canine mammary gland tumour cell lines

The prevalence of cancer in animals has increased significantly over the years. Mammary tumours are the most common neoplasia in dogs, in which around 50% are presented in the malignant form. Hence, the development and characterization of in vitro models for the study of canine tumours are important for the improvement of cancer diagnosis and treatment. Thus, the aim of this study was to characterize cell lines derived from canine mammary gland neoplasias which could be further used for basic and applied oncology research. Samples of canine mammary carcinomas were taken for cell culture and 2 cell lines were established and characterized in terms of cell morphology, tumourigenicity and global gene expression. Both cell lines presented spindle‐shape morphology and shown common malignant features as in vitro invasion potential and expression of epithelial and mesenchymal proteins. Also, we found gene expression patterns between the 2 cell cultures in comparison to the normal mammary gland tissue. Cells from M25 culture showed a higher invasion and in vivo tumourigenic potential, associated to the overexpression of genes involved in focal adhesion and extracellular matrix communication, such as FN1, ITGA8 and THBS2. The phenotypic characterization of these cells along with their global gene expression profile potentially determine new therapeutic targets for mammary tumours.     

Influence of E‐cadherin genetic variation in canine mammary tumour risk,clinicopathological features and prognosis

E‐cadherin is a cell adhesion molecule that participates in several cellular processes that guarantee the maintenance of structural and functional integrity of epithelial tissues. E‐cadherin plays an important role in mammary carcinogenesis, and various studies have demonstrated the effect of CDH1 genetic variation in risk, progression and biological behaviour of human breast cancer. Although there are some recognized genetic variations in canine CDH1 gene, their influence in canine mammary tumour development and progression has not been previously evaluated. In this study, we aim to assess the influence of CDH1 SNPs rs850805755, rs852280880 and rs852639930 in the risk, clinicopathological features and clinical outcome of canine mammary tumours. A case‐control study was conducted involving 206 bitches with mammary tumours and 161 bitches free of mammary neoplasia. CDH1 SNPs rs850805755 and rs852280880 were associated with a decreased risk and a later onset of mammary tumour development. Furthermore, these SNPs were related to the development of small size carcinomas, of low histological grade and low nuclear pleomorphism. SNP rs852639930 was associated with the development of small size tumours with a non‐infiltrative, non‐invasive growth pattern. Data from the present investigation demonstrate that these CDH1 genetic variants could have a protective role in canine mammary tumours, by being associated with low risk of tumour development, delayed onset of the disease and less aggressive clinicopathological features.     

Expression of genes related to liver fatty acid metabolism in fat‐tailed and thin‐tailed lambs during negative and positive energy balances

Fat‐tailed sheep breeds can tolerate periods of negative energy balance without suffering from elevated concentration of plasma non‐esterified fatty acid (NEFA). This ability was attributed to unique metabolism of fat‐tailed adipose depot, whereas role of liver as an influential organ in fatty acid metabolism was not evaluated yet. Hence, current study was conducted to evaluate the effects of negative and positive energy balances on liver expression of genes related to fatty acid metabolism in fat‐tailed and thin‐tailed lambs. Lambs experienced negative (21 days) and positive (21 days) energy balances and were slaughtered at the beginning and end of negative energy balance and at the end of positive energy balance. Real‐time quantitative polymerase chain reaction (RT‐Q‐PCR) was conducted to evaluate changes in gene expression. Expression of diglyceride acyltransferase 1 (DGAT1), 3‐hydroxy‐3‐methylglutaryl‐CoA synthase 2 (HMGCS2) and apolipoprotein B (APOB) was not affected by genotype, energy balance and their interaction. Expression of carnitine palmitoyltransferase 1 (CPT1) was significantly higher in liver of fat‐tailed comparing to thin‐tailed lambs regardless of energy balance (p < 0.02). Catalase mRNA abundance was increased in response to negative energy balance (p < 0.02), and severity of this enhancement was higher in fat‐tailed lambs (p < 0.06). Expression of CPT1 was positively correlated with expression of HMGCS2 in both fat‐tailed (p < 0.05) and thin‐tailed lambs (p < 0.002); however, the correlation was weaker in fat‐tailed lambs (0.72 vs. 0.57, respectively, for thin‐tailed and fat‐tailed lambs). There was a positive correlation between DGAT1 and APOB genes expression in fat‐tailed lambs (0.94; p < 0.001), whereas this correlation was not observed in thin‐tailed lambs. Results demonstrate that liver of fat‐tailed lambs has higher capacity for metabolism of mobilized NEFA exposed to liver during negative energy balance.     

Effect of supplemental β‐carotene compared to retinyl palmitate on fatty acid profile and expression of mRNA from genes involved in vitamin A metabolism in beef feedlot cattle

To examine the effects of dietary β‐carotene (βC) or retinyl palmitate (RP) on fatty acid (FA) profile and mRNA expression, samples were collected from 24 Angus‐cross calves that were allotted to four treatments consisting of RP supplemented at 2200 IU/kg, and synthetic β‐carotene (SβC) supplemented at one, five or 10 times RP. Longissimus muscle (LM) cis‐9, trans‐11 conjugated linoleic acid was greater in RP compared to SβC1X (P = 0.04). The polyunsaturated:saturated FA increased linearly (P = 0.04) in the LM as dietary SβC increased. Expression of βC oxygenase 2 (βCO2), an enzyme that cleaves β‐carotene, was greater in the LM for SβC1X compared to RP and decreased linearly as SβC increased (P ≤ 0.02). Peroxisome proliferator activated receptor γ (PPARγ) expression in the LM increased in SβC1X compared to RP (P = 0.03); however, PPARγ and retinoic acid X receptor α (RXRα) expression decreased linearly (P = 0.02) in the LM with increasing SβC. Retinoic acid receptor α (RARα) expression tended (P = 0.10) to decrease linearly in the LM with increased SβC. In conclusion, SβC supplementation increased mRNA expression of some lipogenic genes in the LM, but increasing dietary SβC inhibited their expression and tended to increase polyunsaturated FA.