The Mechanism Of Aerobic Glycolysis In Chloroquine Induced Death Of Colon Cancer Cells

BackgroundColon cancer is the most common gastrointestinal malignancies,and the mortality rate of colorectal cancer in China ranks fifth among all types of cancer.Tumor cells still get energy in the form of glycolytic enzymes when oxygen is abundant,an effect known as the "Warburg" effect,also known as aerosols.Studies have shown that aerobic glycolysis plays a very important role in tumor development,but the role of aerobic glycolysis with colorectal cancer is rarely reported.Chloroquine(CQ)is a widely spoken antimalarial drug.At the same time,chloroquine has also played a good role in the treatment of rheumatoid arthritis,systemic lupus erythematosus and nodule disease and other acute and chronic inflammation.The study found that in tumor cells,chloroquine can also induce autophagy and induce apoptosis by affecting the cell cycle,and has good anti-tumor potential.However,the relationship between chloroquine and colorectal cancer treatment is not yet known.ObjectivesTostudy the effect of chloroquine on the biological behavior of colon cancer cells,to clarify the relationship between chloroquine causing the death of colorectal cancer cells and aerobic glycolytic enzymes,and to provide new drugs for clinical treatment of colon cancer.Methods1.CCK8 experiments detected the effects of CQ on the survival and proliferation of colorectal cancer cells RKO,HCT-116,LOVO,CACO2.2.Flow cytometry detects the apoptosis of CQ to colorectal cancer cells RKO and HCT-116.3.WB detects the effect of CQ on colorectal cancer cells RKO,HCT-116 apoptosisrelated proteins.4.The effect of CQ on the levels of rectal cancer cells RKO,HCT-116 aerobic glycolysis was detected using glucose intake kits and lactic acid test kits.5.The effects of CQ on glycolyzyme-related kinase HK2 m RNA and protein expression in colorectal cancer cells were detected using real-time quantitative chain reactions(real-time quantitative PCR,qRT-PCR)and WB.6.The expression level of HK2 in colorectal cancer cells RKO and HCT-116 was reduced by RNA interference technology,and protein and m RNA levels were verified using WB and qRT-PCR.7.The effect of CQ on the levels of rectal cancer cells RKO,HCT-116 aerobic glycolysis was detected by using glucose intake kits and lactic acid test kits to detect interference with HK2.8.CCK8 experiments tested the effects of CQ-combined HK2 sh RNA and its inhibitors on the survival of colorectal cancer cells RKO and HCT-116.9.WB detects the effects of CQ-combined HK2 sh RNA and its inhibitors on colorectal cancer cells RKO,HCT-116 apoptosis-related proteins.Results1.CCK8 results showed that with the increase of CQ concentration,there was a significant inhibition effect on the survival and proliferation of colorectal cancer cells RKO,HCT-116,LOVO,CACO2.The difference is statistically significant(P<0.05).2.The results of flow cytometry showed that the concentration of CQ increased,and the apoptosis rate of colorectal cancer cells RKO and HCT-116 also increased gradually.3.WB results showed an increase in CQ concentrations and a gradual increase in the expression of the apoptosis-related protein Claved-caspase-3 in colorectal cancer cells RKO and HCT-116.The difference is statistically significant(P<0.05).4.Glucose intake detection and lactic acid testing results showed that colorectal cancer cells RKO,HCT-116 after CQ treatment of glucose intake increased,lactic acid production increased.The difference is statistically significant(P<0.05).5.qRT-PCR and WB results showed that after CQ treatment,HK2 expression at m RNA levels changed and protein expression increased.6.qRT-PCR and WB verified HK2 interference efficiency,and the results showed that HD2 decreased in m RNA and protein level expression after interference.The difference is statistically significant(P<0.05).7.Glucose intake test and lactic acid test results showed that CQ reduced colorectal cancer cells RKO,HCT-116 glucose intake and lactic acid production after interference with HK2.The difference is statistically significant(P<0.05).8.CCK8 and WB results show that CQ joint sh HK2 can further inhibit the survival of colorectal cancer cells RKO,HCT-116,and promote the apoptosis-related protein Clavedcaspase-3 upward.Conclusions1.CQ inhibits the survival and proliferation of colorectal cancer cells RKO,HCT-116,LOVO,CACO2,and induces its apoptosis.2.CQ increases the level of colon cancer cells RKO,HCT-116 aerobic glycolysis.3.Inhibition of aerobic enzymes can further promote the apoptosis of colorectal cancer cells induced by CQ.