Previous reports showed that activation of the PI3K/mTOR pathway is associated with chemoresistance in ovarian cancer cells, and this can be reversed by combining chemotherapy with the mTORC1/2 inhibitor vistusertib (AZD2014). This combination is now being pursued clinically. In this study, we used NMR spectroscopy to establish biomarkers for this combination in A2780Cis human ovarian carcinoma cells. We report that this combination resulted in decreases in lactate, PC, GPC and tCho and was associated with inhibition of glycolysis- and choline-regulatory enzymes. This indicates that choline metabolites and lactate may provide potential non-invasive biomarkers of tumour response to this combination therapy.