Non-melanoma skin cancer (NMSC) is the most common type of cancer worldwide. Treatment options for NMSC include Photodynamic therapy (PDT). PDT is used mainly for small and superficial tumors, including actinic keratosis, in situ squamous cell carcinoma and basal cell carcinoma. The principal compounds used in PDT for NMSC are ALA and MAL, precursors of the photoactive compound protoporphyrin IX (PpIX). PDT produces very satisfactory results in clinic; however, some cells can survive, producing tumor relapses and/or increases in their clinical and biologic aggressiveness. Even intrinsic cell tumor factors (specific mutations), tumor microenvironment, constituted by cancer cells, immune and endothelial cells, extracellular matrix fibers and particularly fibroblasts (cancer-associated fibroblasts, CAFs), play a crucial role in modulation the activity, growth, and the resistance to cancer therapies. Therefore, our objective is to define factors implicated in the response to PDT with the goal of improving PDT by itself or by combining it with other treatment modalities to target both, cancer cells and CAFs, to eradicate the tumor lesion. In this sense, we have evaluated the combination of PDT with 5-FU, metformin and rapamycin in bidimensional and tridimensional in vitro models (spheroids) as well as in xenografts.