The onset of resistance to cisplatin still limits its use in the chemotherapy of ovarian cancer and, despite several mechanisms of resistance have been discovered, they are not exhaustive. The aim of this study was to identify which other pathways are exploited by cancer cells to escape cisplatin cytotoxicity, to possibly prevent or overcome the phenomenon with new pharmacological approaches. The increase of anaerobic glycolysis, even in the presence of oxygen (Warburg effect), is the first observation indicating the alteration of energetic metabolism used by tumor cells as a strategy to adapt and grow independently from the availability of the substrate. This evidence suggested us to investigate the hypothesis that a similar metabolic strategy might be of relevance in resistance to cisplatin. Recently it has been shown that only approximately 1% of intracellular platinum is bound to nuclear DNA, while the great majority of the intracellular drug is available to interact with other nucleophilic sites including but not limited to phospholipids, cytosolic, cytoskeletal and membrane proteins, RNA and mitochondrial DNA. mtDNA, unlike nDNA, does not possess efficient repair systems and is therefore more susceptible to the onset of mutations often associated to cancer development, loss of tumor suppressor, activation of oncogenes and mitochondrial dysfunctions often related with an increase of glycolytic activity. Therefore, the aim of this study was to investigate the energetic metabolism and the mitochondrial function of cisplatin-resistant (C13) and sensitive (2008) ovarian cancer cells with different experimental approaches. Results revealed that resistant cells present a significant reduced respiratory chain activity correlated to a lower mitochondrial mass, altered mitochondrial morphology as well as a metabolomic profile typical of a lipogenic phenotype. To investigate the role of mtDNA and nDNA in the mitochondrial and metabolic remodeling of cisplatin-resistant line, cancer cells (2008-C13) were used to generate transmitochondrial hybrids (H2008-HC13). Mitochondrial DNA of parental and hybrid cells was sequenced, showing similar, almost non pathological, polymorphisms. Interestingly, investigating the energetic metabolism and the mitochondrial structure of hybrids, no differences were observed between H2008 and HC13. These data demonstrated that the metabolic reprogramming of C13 cells was not dependent from mtDNA, but was controlled by nuclear factors. Having regard to these data, the activity of some nuclear transcription factors (HIF-1α, and c-Myc) involved in the metabolic reprogramming of tumor cells, has been evaluated and it has been highlighted a different expression of some of their target genes involved in the glycolytic flux. Finally, the metabolic profile of 2008-C13 cells has been outlined by LC-MS which evidenced some interesting differences in aminoacids, phospholipids and antioxidants content