Phenotypic screening of a library of compounds against metastatic and non-metastatic clones of a canine mammary gland tumour cell line

- Metastases are related to a poor prognosis for canine mammary gland tumours (CMGTs).
- Phenotypic compound screening was conducted to identify agents selective for a metastatic CMGT clone.
- Mitochondrial respiratory chain complex inhibitors (MRCIs) were identified as candidate agents.
- MRCIs showed selective growth inhibitory effect on the metastatic CMGT clone.
- MRCIs exerted their anti-tumour activity through an AMPK-dependent or -independent mechanism.

Metastases are associated with a poor prognosis for canine mammary gland tumours (CMGTs). Metastatic and non-metastatic clones were isolated previously from a single malignant CMGT cell line. The difference in metastatic potential between the two cell lines was hypothesised to be associated with distinct cellular signalling. The aim of this study was to screen for compounds that specifically target metastatic cells in order to improve CMGT therapeutic outcomes. The two clonal cell lines were characterised by transcriptome analysis and their sensitivity to a library of 291 different compounds was compared.The metastatic clone exhibited elevated expression of molecules associated with degradation of the extracellular matrix, epithelial-mesenchymal transition and cancer stem cell phenotype. This was confirmed using a matrigel invasion assay and by assessment of aldehyde dehydrogenase activity. The mitochondrial respiratory chain complex inhibitors (MRCIs; rotenone, antimycin and oligomycin) significantly inhibited the growth of the metastatic clone. Although MRCIs similarly depleted mitochondrial ATP in both clones, the subsequent cellular response was different, with toxicity to the metastatic clone being independent of AMP-activated protein kinase activity. The results of this study suggest a potential utility of MRCIs as anti-tumour agents against metastatic CMGTs. Further studies are needed to investigate the clinical utility of MRCIs and to determine the association between MRCI sensitivity and malignancy.