IADR Abstract Archives
Characterization of Mitochondrial Dysfunction and Biogenesis in Oral Cancer's Micro-environment
Objectives: Cancer is primarily a metabolic disease involving disturbances in cellular energy metabolism.Based on“Reverse Warburg Effect”,we try to develope a novel co-culture system to compare mitochondrial function and energy production between normal oral fibroblasts(NOFs) and cancer associated fibroblasts (CAFs),and to study their pathophysiological significance in micro-environment of oral squamous cell carcinoma (OSCC).
Methods: Cafs and Nofs were both monolayer cultured,and co-cultured respectively with normal oral keratinocytes(NOKs) and OSCC.Stained and analysed them by luminescent reader,flow cytometry,confocal microscope and western blot for ATP production,mitochondrial volume,mitochondrial membrane potential(MMP),reactive oxygen species (ROS),Lactate,mitochondrial morphology,caveolin-1 and monocarboxylate tranporter 4(MCT-4).Mean and standard deviation were plotted for 3 replicates from each condition.
Results: After co-cultured with OSCC,CAFs showed decreased ATP production,ROS,Caveolin-1 expression and lower MMP,but mitochondrial volume and MCT-4 expression were increased.Interestingly,NOFs showed increased ATP production,ROS,lactate,MCT-4 expression,but decreased MMP,mitochondrial volume and Caveolin-1 expression after co-cultured with OSCC.When co-cultured with NOKs,both CAFs and NOFs didn’t show significant difference compared with monolayer culture. Confocal work for NOFs-NOKs and NOFs-OSCC co-culture system showed that mitochondrial tranfer more easily happen and unidirection from NOFs to OSCC.
Conclusions: OSCC cells can induce the“Reverse Warburg Effect”in two kinds of stromal fibroblasts—NOFs and CAFs—which can present mitochondrial dysfunction and oxidative stress in differernt ways that fuel the ATP production in OSCC and drive their mutagenic evolution towards a more aggressive phenotype.Interestingly,we observed that metabolic coupling and mitochondrial transfer occured between stromal fibroblasts and OSCC.As the precursor of CAFs,NOFs play a more significant role in invasive tumor front(ITF) than CAFs,which dramatically increase aggressive behavior of OSCC and associate with early tumor ecurrence,metastasis and poor clinical outcome.
Methods: Cafs and Nofs were both monolayer cultured,and co-cultured respectively with normal oral keratinocytes(NOKs) and OSCC.Stained and analysed them by luminescent reader,flow cytometry,confocal microscope and western blot for ATP production,mitochondrial volume,mitochondrial membrane potential(MMP),reactive oxygen species (ROS),Lactate,mitochondrial morphology,caveolin-1 and monocarboxylate tranporter 4(MCT-4).Mean and standard deviation were plotted for 3 replicates from each condition.
Results: After co-cultured with OSCC,CAFs showed decreased ATP production,ROS,Caveolin-1 expression and lower MMP,but mitochondrial volume and MCT-4 expression were increased.Interestingly,NOFs showed increased ATP production,ROS,lactate,MCT-4 expression,but decreased MMP,mitochondrial volume and Caveolin-1 expression after co-cultured with OSCC.When co-cultured with NOKs,both CAFs and NOFs didn’t show significant difference compared with monolayer culture. Confocal work for NOFs-NOKs and NOFs-OSCC co-culture system showed that mitochondrial tranfer more easily happen and unidirection from NOFs to OSCC.
Conclusions: OSCC cells can induce the“Reverse Warburg Effect”in two kinds of stromal fibroblasts—NOFs and CAFs—which can present mitochondrial dysfunction and oxidative stress in differernt ways that fuel the ATP production in OSCC and drive their mutagenic evolution towards a more aggressive phenotype.Interestingly,we observed that metabolic coupling and mitochondrial transfer occured between stromal fibroblasts and OSCC.As the precursor of CAFs,NOFs play a more significant role in invasive tumor front(ITF) than CAFs,which dramatically increase aggressive behavior of OSCC and associate with early tumor ecurrence,metastasis and poor clinical outcome.