The Effects Of Glucose Deprivation For HeLa Cell Activity And Cell Cycle

The occurrence of malignant tumor is associated with abnormal cellular metabolism.Tumor cells show an increased rate of glucose uptake,glycolysis metabolism and accumulation of extracellular lactic.Glycolysis provides tumor cells with enough ATP.The enough ATP is advantageous to proliferation and invasion of tumor cells.The glycolytic pathway shows low efficiency of productivity compared with the oxidative phosphorylation pathway.Glucose uptake from the extracellular environment in glycolytic pathway is roughly ten times more than normal cells.Because of tumor cells need absorb a lot of glucose,People pay more attention to the method of cancer treatment with glucose deprivation.In this study,we investigated HeLa cells and umbilical cord normal cells treated with glucose deprivation to analysis the influence of cells activity and proliferation,preliminary clarify the effects of glucose deprivation for HeLa cells cycle molecular mechanism,which will provide theoretical basis for starvation treatment of cancer.The results are listed as follows:1.With glucose deprivation separately working on HeLa cells and umbilical cord normal cells,glucose deprivation working for 24 h,48 h,72 h with 1mmol/L and 0 mmol/L glucose,the survival rate was determined by MTT assay.The results showed that,with 1 mmol/L glucose deprivation working on HeLa cells,cells inhibitory rate was 34.6%?57.14%and 73.09%,with 1 mmol/L glucose deprivation working on umbilical cord normal cells,cells inhibitory rate was 0.36%?26.4%and 66.3%;with 0 mmol/L glucose deprivation working on HeLa cells,cells inhibitory rate was 46.84%?74.09%and 96.94%,with 0 mmol/L glucose deprivation working on umbilical cord normal cells,cells inhibitory rate was 7.61%?58.6%and 82.1%.Compared with the umbilical cord normal cells,with glucose deprivation,the inhibition activity of HeLa cells more obvious.Cell morphology was observed under the inverted microscope,trypan blue staining to observe cell survival rate.The results showed that:with 0 mmol/L glucose deprivation working,most cells apoptosis after 24 h in HeLa cells,most cells apoptosis after 48 h in umbilical cord normal cells;with 1 mmol/L glucose deprivation working,most cells apoptosis after 48 h in HeLa cells,most cells apoptosis after 72 h in umbilical cord normal cells.Compared with the umbilical cord normal cells,with glucose deprivation,HeLa cells are more prone to apoptosis than umbilical normal cells.2.In order to better simulate the human body environment,investigate the effects of glucose deprivation on tumor cells and normal cells,co-cultivate umbilical cord normal cells and HeLa cells.With glucose deprivation together working on HeLa cells and umbilical cord normal cells,the results are consistent with separate cultivate.with 0 mmol/L glucose deprivation working on co-cultivate cells.In HeLa cell groups,most of the cells in the middle of apoptosis state after 24 h,almost all cells is in a state of late apoptosis after 48 h,all cells in a state of late apoptosis after 72 h.In umbilical cord normal cell groups,a few cells in the middle of apoptosis state after 24 h,most of the cells in the middle of apoptosis state after 48 h,almost all in a state of late apoptosis after 72 h.With 1 mmol/L glucose deprivation working on co-cultivate cells.In HeLa cell groups,a few cells in the middle of apoptosis state after 24 h,most of the cells in the middle of apoptosis state after 48 h,almost all in a state of late apoptosis after 72 h.In umbilical cord normal cell groups,keep normal growth after 24 h,a few cells in the middle of apoptosis state after 48 h,most of the cells in the middle of apoptosis state after 72 h.With glucose deprivation,HeLa cells are more prone to apoptosis than umbilical cord normal cells under the co-cultivate conditions.3.Mitochondria activity was observed by Mito-Tracker Green,flow cytometry instrument to detect the cell cycle,Western Blot detected expression changes in cell cycle proteins.With glucose deprivation working on HeLa cells for 48 h,the mitochondria activity was enhanced significantly,the cell cycle stagnation in the G1 phase,the decreased expression of CyclinA?CyclinD?CyclinE?CyclinH,the decreased expression of CDK2?CDK4?CDK6?CDK9 and the increased expression of CDK7,the decreased expression of P16?P18?P19?P21?P53 and the increased expression of P15 and P27.The others cell cycle related proteins PCNA and MCM was decreased expression.With glucose deprivation on HeLa cells,cell shrinkage,the mitochondria activity was enhanced,perhaps improve glucose utilization,provide more energy for cells growth.Glucose deprivation can obviously inhibit the cell proliferation and cell cycle process of HeLa cells.the cells stagnation in the G1 phase,the mechanism of cell cycle change might be through the cell cycle dependency protein kinase inhibitors of p27 protein increased expression,inhibiting the activity of CyclinE-CDK2 complex,make the cell cannot pass the G1/S transition,thus inhibiting cell proliferation and promoting cell apoptosis.With glucose deprivation,HeLa cells are more prone to apoptosis than umbilical normal cells.Glucose deprivation can obviously inhibit the growth of HeLa cells,Change the cell morphology,inhibition the cell cycle,the cells stagnation in the G1 phase.The mechanism might be through the cell cycle dependency protein kinase inhibitors of p27 protein increased expression,inhibiting cell proliferation and promoting cell apoptosis.