Effect Of Celastrol On Proliferation And Aerobic Glycolysis Of Gastric Cancer Cells

Objective The study investigated the effects of celastrol on the proliferation and aerobic glycolysis of gastric cancer cell line BGC-823,and explored whether celastrol can regulate aerobic glycolysis and inhibit the proliferation of BGC-823.Methods After treatment of celastrol,BGC-823 cells viability was detected by MTT assay,cell proliferation ability was detected by colony formation assay.Cell apoptosis and cell cycle distribution was detected by flow cytometry,and cell growth and morphology were observed under light microscope.Glucose utilization,lactic acid production,hexokinase(HK) activity and lactate dehydrogenase(LDH) activity was detected by spectorphotometry.Western blot was conducted to detect the different expression of GLUT1,HKII,PKM2 and LDHA between BGC-823 cells and GES cells,then the expression of the above protein was detected in BGC-823,which was treated with experimental concentration of celastrol,by western blot.Results1.BGC-823 cells were treated with gradient concentration of celastrol for24 h,48 h,and 72 h,MTT assay showed that the cell viability of the drug-added group decreased gradually compared with the control group.The IC50 values of BGC-823 cells treated with celastrol were(1.66±0.05)?mol/L,(1.49±0.07)?mol/L,(1.32±0.10)?mol/L at 24 h,48 h and 72 h,respectively(N=3,(?)±SD).2.After treatment of celastrol on BGC-823 cells,the plate clone formation assay showed that the cell clone formation rate of each drug-added group were(57.63±5.78)%,(32.64±3.27)%,(0.38±0.27)%,respectively,which were appreciably lower than that of the control group,which was(77.60 ± 6.41)%(P<0.01),and the inhibition is concentration dependent.3.After treatment of celastrol on BGC-823 cells,cells were observed under the inverted microscope,we found that,with the increase of drug concentration,the number of viable cells in the culture dish was reduced,the number of suspended cells was increased,the cell density decreased,and the cell morphology changed from elliptical to fusiform.4.After treatment of celastrol on BGC-823 cells,the apoptosis rate of the control group and each experimental group were(14.90±0.87)%,(18.12±0.50)%,(19.15±0.04)%,(24.15±1.76)%.Compared with the control group,the apoptosis rate was increased(P<0.05).And the apoptosis rate was concentration dependent.5.After treatment of celastol on BGC-823 cells,cell division were blocked,which blocking at G2/M phase.6.Celastrol inhibited the utilization of glucose(P<0.05) and the production of lactic acid(P<0.01) in the aerobic glycolysis of BGC-823 cells,and the inhibition is concentration dependent.7.In gastric cancer cells,the protein expression of GLUT1,HK?,LDHA was increased(P<0.05),and the expression of PKM2 was decreased(P<0.05),compared with gastric epithelial cells.In addition,Celastrol can inhibit the protein expression of GLUT1,HK?,LDHA,which play an important role in the aerobic glycolysis of BGC-823 cells(P<0.05).Celastrol has no affection on the expression of PKM2(P>0.05).8.The inhibitory effect of celastrol on the activity of key enzymes in aerobic glycolysis of BGC-823 cells was not significant.HK activity was inhibited at a concentration of 1.5 ?mol/L L(P<0.05),and LDH activity was inhibited at a concentration of 1.5 ?mol/L(P<0.05).Conclusions In gastric cancer cell line BGC-823,celastrol can directly inhibit the key proteins GLUT1,HK? and LDHA in aerobic glycolysis pathway,and inhibit the activity of HK and LDH enzyme at high concentration,thus celastrol can hinder the ingestion of glucose and the production of lactic acid,and inhibit the aerobic glycolysis.This reduces the supply of cellular energy and the raw materials required for the synthesis of biomacromolecules in BGC-823,which hindering cell cycle progression,promoting apoptosis,and inhibiting the growth and proliferation of BGC-823.