Mithun Kailavasan1, Steven Reynolds1, Adriana Bucur1, Samira Kazan2, Tooba Alizadeh2, Gillian M. Tozer2, Ishtiaq Rehman3, Martyn Paley1
1Academic Radiology, University of Sheffield, Sheffield, Yorkshire, United Kingdom; 2Oncology, University of Sheffield, Sheffield, Yorkshire, United Kingdom; 3Human Metabolism, University of Sheffield, Sheffield, Yorkshire, United Kingdom
Prostate cancer cells with varying metastatic potential (LNCaP and LNCaP-LN3) were cultured in a customized cell bioreactor system, adapted for a 9.4T Nuclear Magnetic Resonance (NMR) probe, which allowed extended cell survival. Several metabolite levels were continuously measured over time, including lactate, fatty acid, alanine, choline, glutamine and creatine, both with and without the addition of a pyruvate dehydrogenase kinase inhibitor, dichloroacetate (DCA). Zymography was used to assess LDH isoenzymes. Rat p22 sarcoma cells were used as a control. Zymography assays (n=4), showed an absence of the LDH-B subunit in both LNCaP-LN3 and rat p22 cell lines. LNCaP-LN3 and rat p22 cells had a 18600mol/108 cells (p<0.001) and 6600mol/108 cells (p=0.039) median difference, respectively, in lactate levels compared to LNCaP. Median Cho/Cr ratios at 1hr decreased following DCA treatment in LNCaP and LNCAP-LN3 (p<0.001). Creatine levels increased over time in all 3 cell lines by showing efficiency of the bioreactor design.