The Study Of Energy Metabolism In Bladder Cancer Cells In Co-culture Conditions Using A Microfluidic Chip

Objective: Systematically analyze the change of mitochondrial-related proteins of bladder cancer cells and tumor-associated fibroblasts. To investigate the characteristics of bladder cancer cell energy metabolism.Method:(1) Building a co-culture system of bladder tumor cells and fibroblasts with microfluidic chip, a three-dimensional culture technique.(2) Determining and calculating the concentration of lactic acid in the medium of different groups using automatic microplate reader(3) Qualitative analysis of mitochondria-related proteins by immunofluorescence staining.(4) Quantitative analysis of mitochondrial associated protein by Western blot.(4) Using SPSS software for data analysis.Result:(1) Determination of lactic acid concentration: the result in descending order is the experimental group, the control group of T24 cell, the control group of fibroblasts.(2) Qualitative results: In the control group, the mitochondrial-related proteins fluorescence intensity of the fibroblasts was higher than of the cancer cells and the fluorescence intensity of the fibroblasts was lower in the experimental group. The mitochondrial-related proteins fluorescence intensity of the cancer cells was higher in the experimental group than in the control group, and the fibroblasts shared the performance of the opposite.(3) Quantitative results: expression of mitochondria-related proteins of fibroblasts was higher than of cancer cells in the control group, and showed the exact opposite results in the experimental group(p<0.05); mitochondria-related proteins expression of cancer cells in experimental group was higher than in control group, and the fibroblasts sharehe exact opposite results( p<0.05).Conclusion: The energy metabolism of bladder tumor cells does not parallele the ‘Warburg effect’ as ‘even under the condition of sufficient oxygen, cancer cells are still function in the form of glycolysis’. Bladder cancer cells also have efficient oxidative phosphorylation process: cancer cells promote the glycolysis of the adjacent interstitial cells, causing increased formation of nutritional precursor. These high-energy metabolites are directional transit transferred to adjacent tumor cells and enter the Krebs cycle. Finally the oxidative phosphorylation is increased and plenty of ATP is produced.