Figure 6. Effect of Rh2E2 on the network-based metabolic reprograming of LLC-1 cancer cells.
A. Rh2E2 decreased the ATP production in LLC-1 cancer cells. B. Rh2E2 enhanced ATP generation in CCD19Lu normal cells. Equal numbers of medium control or Rh2E2-treated cells (LLC-1 or CCD19Lu) were harvested for ATP, ADP and AMP determination by UPLC-MS quantitation with their reference control. The amount of energy metabolites was calculated as pmol/106 cells. C. Schematic diagram of network-based metabolic intervention of Rh2E2 in LLC-1 lung cancer cells. Mitochondrial proteins from Rh2E2-treated LLC-1 cancer cells or CCD19Lu normal cells were harvested for proteomic analysis using iTRAQ. Three biological replicates were compared and analyzed to reduce the variation caused by random biological effects. D. Rh2E2 specifically reduced the production of acetyl CoA in LLC-1 lung cancer cells. E. Immunoblotting validation of the identified metabolic enzymes in TCA cycle. F. Rh2E2 enhanced the accumulation of α-ketoglutarate, the energy metabolite of TCA cycle.