Background: Circulating tumor cells (CTC) have relevance as prognostic markers in breast cancer. However, the functional properties of CTCs or their molecular characterization have not been well-studied. Experimental models indicate that only a few cells can survive in the circulation and eventually metastasize. Thus, it is essential to identify these surviving cells capable of forming such metastases. Methods: We isolated viable CTCs from 50 peripheral blood samples obtained from 35 patients with advanced metastatic breast cancer using RosetteSepTM for ex vivo culture. The CTCs were seeded and monitored on plates under low adherence conditions and with media supplemented with growth factors and Nanoemulsions. Phenotypic analysis was performed by immunofluorescence and gene expression analysis using RT-PCR and CTCs counting by the Cellsearch® system. Results: We found that in 75% of samples the CTC cultures lasted more than 23 days, predicting a shorter Progression-Free Survival in these patients, independently of having ≥5 CTC by Cellsearch®. We also observed that CTCs before and after culture showed a different gene expression profile. Conclusions: the cultivability of CTCs is a predictive factor. Furthermore, the subset of cells capable of growing ex vivo show stem or mesenchymal features and may represent the CTC population with metastatic potential in vivo.

Colour polymorphism is a widespread phenomenon in natural populations of several species. In particular, it is especially common on marine gastropod species from the genus Littorina. Recently, it has been argued that intrapopulation shell colour polymorphism in Littorina fabalis could be caused by negative frequency-dependent sexual selection via a mechanism of mate choice (indirectly estimated via negative assortative mating). Here we try to determine the existence of negative assortative mating in three species of the subgenus Neritrema (L. fabalis, L. obtusata, L. saxatilis) that share a similar shell colour polymorphism, in order to ascertain if this mechanism could represent an ancestral character in this subgenus that could be contributing to the maintenance of the colour polymorphism in each species.Here, we collected or reanalysed from previous studies a sample of mating pairs of the three species from seven locations from NW Spain and NE Russia and estimated assortative mating using the I PSI index. Our results show that all species and populations show a systematic tendency towards negative assortative mating when shell colour is grouped in the broad categories: 'light' and 'dark'. Although, a more detailed analysis of each colour individually suggests that shell colour may not be the main target of assortative mating, but perhaps physically linked to another trait or through pleiotropic effects. This hypothesis opens interesting new lines of research in Littorina snails.

CTCs have extensively been used for the monitoring and characterization of metastatic prostate cancer, but their application in the clinic is still very scarce. Besides, the resistance mechanisms linked to prostate cancer treatment remain unclear. Liquid biopsies represent the most promising alternative due to the complexity of biopsying bone metastasis and the duration of the disease. We performed a prospective longitudinal study in CTCs from 20 castration-resistant prostate cancer patients treated with docetaxel. For that, we used CellSearch® technology and a custom gene expression panel with qRT-PCR using a CTCs negative enrichment approach. We found that CTCs showed a hybrid phenotype during the disease, where epithelial features were associated with the presence of ≥ 5 CTCs/7.5 mL of blood, while high relative expression of the gene MYCL was observed preferentially in the set of samples with < 5 CTCs/7.5 mL of blood. At baseline, patients whose CTCs had stem or hybrid features showed a later progression. After 1 cycle of docetaxel, high relative expression of ZEB1 indicated worse outcome, while KRT19 and KLK3 high expression could predisposed the patients to a worse prognosis at clinical progression. In the present work we describe biomarkers with clinical relevance for the prediction of early response or resistance in castration-resistant prostate cancer patients. Besides, we question the utility of targeted isolated CTCs and the use of a limited number of markers to define the CTCs population.

Colour polymorphism is a widespread phenomenon in natural populations of several species. In particular, it is especially common on marine gastropod species from the genus Littorina. Recently, it has been argued that intrapopulation shell colour polymorphism in Littorina fabalis could be caused by negative frequency-dependent sexual selection via a mechanism of mate choice (indirectly estimated via negative assortative mating). Here we try to determine the existence of negative assortative mating in three species of the subgenus Neritrema (L. fabalis, L. obtusata, L. saxatilis) that share a similar shell colour polymorphism, in order to ascertain if this mechanism could represent an ancestral character in this subgenus that could be contributing to the maintenance of the colour polymorphism in each species. Here, we collected or reanalysed from previous studies a sample of mating pairs of the three species from seven locations from NW Spain and NE Russia and estimated assortative mating using the IPSI index. Our results show that all species and populations show a systematic tendency towards negative assortative mating when shell colour is grouped in the broad categories: ‘light’ and ‘dark’. Although, a more detailed analysis of each colour individually suggests that shell colour may not be the main target of assortative mating, but perhaps physically linked to another trait or through pleiotropic effects. This hypothesis opens interesting new lines of research in Littorina snails.