Effect Of Knocking Down G6PDH On The Biological Characteristics Of Hepatocellular Carcinoma Cells

ObjectiveCells all tend to produce glucose in aerobic glycolysis that is split by the metabolic pathway of the Pentose Phosphate Pathway(PPP)to increase the metabolic intermediates necessary for de novo synthesis of nucleotides,while glucose The 6-phosphate dehydrogenase is the first rate-limiting enzyme of PPP.To this end,the design of the use of HBV replication cell model HepG2.215 cells by lentivirus infection knock down the G6 PDH gene,explore the effects of G6 PDH on the biological characteristics of liver cancer cells.methodsThe cell strain knocked down by G6 PDH will be successfully infected with lentivirus,and through drug screening,a stably expressed monoclonal will be picked and verified.In the biological characteristics of tumors,in vitro experiments were performed through cell growth curves,cell cycle,and apoptosis.In vivo experiments were performed by inoculating cells subcutaneously to observe the time of tumor formation and tumor size,and glucose was detected by detecting lactic acid and by using seahorse.Oxidative metabolism and glycolytic metabolism to verify the effect of G6 PDH on metabolic pathways.results1.The Western blot was used to detect the expression of G6 PDH in HepG2.215 cell line and Knockdown cell line,and the expression was consistent with the expected size.2.In the characteristics of tumor biologics,cell growth,cycle,and apoptosis in G6 PDH knockdown cell group were significantly different from those in Hep G2.215 cell group(P<0.05);In vivo experiments,G6 PDH knock The low-grade subcutaneous tumorigenesis ability and tumor size were significantly lower than those in the control group.In the oxidative metabolic pathway,the maximum respiratory rate and ATP production G6 PDH were lower than in the control group.In the glycolytic metabolic pathway,there was no significant difference between G6 PDH and the control group.conclusionLentivirus knockout confirmed that G6 PDH is involved in the regulation of hepatitis B virus replication and is a gene that promotes tumor growth and proliferation.