摘要
Abstract
Objective To explore the effects of disturbance of lactic acid efflux, occurred by monocarboxylate transporter-1 (MCT1) translocation on u251 cells in vitro induced by CD147/HAbl8G genetic engineering monoclonal antibody,on energy synthesis, pHi and apoptosis of tumor cells. Methods Low and high dose of CD147/HAbl8G monoclonal aitibody were applied to U251 cell in vitro with control group set. The translocation of MCT1 expression was demonstrated by immunofluorescence staining. Intracellular lactate content was detected by spectrophotometer,and ATP content was measured by bioluminescence method. Phos-phofructokinase (PFK) activity was checked by continuous colorimetry. pHi was recorded by BCKCF-AM fluorescence probe, and apoptopsis rate was detected by flow cytometer. Results Interfered by monoclonal aitibody,the expression of MCT1 was translocated from cell membrane to cytoplasma, intracellular lactate content significantly increased, and ATP content obviously decreased [control group: (0. 831 ± 0. 036) × 10 8 mmol/L, low dose group: (0. 592 ± 0. 047) × 10 8 mmol/L, high dose group: (0. 332 + 0.042)×10 8 mmol/L,P<0. 01]. PFK activity decreased [control group: (0. 855 ± 0. 076)mmol ? Min 1 ?L1 , low dose group: (0. 602± 0. 057)mmol ? Min 1 ? L1 ,high dose group: (0. 382 ± 0. 049)mmol ? Min 1 ? L1 ,P<0. 01]. The difference of pHi was significant among 3 groups [control group: (7. 27 ± 0. 03) , low dose group: (6. 77 ± 0. 04) , high dose group: (6. 31 ± 0. 02) , P< 0. 01]. Apoptosis rate increased [control group: (8. 45 ± 1. 26)%, low dose group: (16. 87 ± 3. 26)%, high dose group: (28. 92 ± 3. 01) ,P<0. 01]. Conclusion The translocation of MCT1 expression can effectively restrain the clearance of intracellular lactate generated by glycolysis,reduce ATP output,acidate cell microenvironment,and promote cell apoptosis.关键词
单羧酸转运蛋白-1/糖酵解/细胞内pH值/细胞凋亡Key words
monocarboxylate transporter-1/ glycolysis /pHi/ apoptopsis
