Imagej western blot quantification drivers#
Imagej western blot quantification driver#
Imagej western blot quantification skin#

Imagej western blot quantification driver#

Notably, the most frequent driver mutations are already present in benign nevi and are not indicative of disease outcome ( 15–17). Melanoma is associated with a high mutation burden due to UV-damage arising from sun exposure.

Imagej western blot quantification skin#

Here, we have searched for novel RBPs implicated in cancer progression and metastasis using melanoma as a model system.Ĭutaneous melanoma is the most aggressive form of skin cancer, and a disease of increasing incidence where mortality remains high ( 14). Despite their prevalence, the number of RBPs with established roles in tumorigenesis and metastatic progression is small, and their molecular mechanisms of action are poorly understood. An interesting group are enzymes that have dual roles as RBPs and may connect cellular metabolism with RNA regulation ( 11–13). The RBP family includes a wide spectrum of entities, from well-known RBPs containing classical RNA-binding domains (RBD) to proteins without any discernible RBD, generally containing intrinsically disordered regions that participate in RNA binding. Recent high-throughput experiments, including RNA-interactome capture (RIC) and other unbiased technologies, have greatly expanded the number of potential members of this family, which are counted in the thousands ( 9, 10). RBPs regulate all aspects of post-transcriptional gene expression, and have been shown to modulate virtually all cancer hallmarks ( 4–8). For this reason, RNA-binding proteins (RBPs) are gaining considerable attention in the cancer field. Epigenetic and post-transcriptional reprogramming allow cancer cells to quickly respond to changing environments and resist a wide variety of stresses. However, non-genetic adaptive mechanisms are increasingly recognized to play fundamental roles in cancer cell plasticity, malignancy and resistance to therapy ( 1–3).

Imagej western blot quantification drivers#

Genetic mutations of oncogenic drivers are at the base of cancer generation. These results exemplify how RIC technologies can be harnessed to uncover novel vulnerabilities of cancer cells. We show that PDIA6 is involved in metastatic progression, map its RNA-binding domain, and find that RNA binding is required for PDIA6 tumorigenic properties. As proof-of-principle we focused on PDIA6, an ER-lumen chaperone that displayed a novel RNA-binding activity.

Extensive functional validation of a selected group of 24 RBPs using five different in vitro assays unveiled unanticipated roles of RBPs in melanoma malignancy. Comparison of RIC profiles of a non-tumoral versus a metastatic cell line revealed prevalent changes in RNA-binding capacities that were not associated with changes in RBP levels. Here, we use RNA interactome capture (RIC) to characterize the melanoma RBPome and uncover novel RBPs involved in melanoma progression. RNA-binding proteins (RBPs) have been relatively overlooked in cancer research despite their contribution to virtually every cancer hallmark.