Based On MicroRNA-1296-mediated MTOR/SLC2A3 Pathway,the Mechanism Of Qingre Huashi Decoction On Gastric Cancer Was Discussed Of Gastric Cancer

Objective: To explore whether mi RNA-1296 can participate in the metabolic reprogramming of gastric cancer by regulating m TOR / SLC2A3 signaling pathway,and further confirm the mechanism of Qingre Huashi Decoction in the prevention and treatment of gastric cancer.Methods: The expression of mi RNA-1296 in 5 human gastric cancer cells(AGS,HGC-27,MGC-803,NCI-N87,MKN-45)was detected by PCR to determine the cells for subsequent experiments.The stable overexpression and silencing of mi RNA-1296 MKN45 cells were constructed by lentivirus transfection technique.The drug-containing serum of Qingre Huashi Decoction was prepared,and the experiment was divided into control group(MKN45),blank serum group,drug-containing serum group,overexpression mi RNA1296,silencing mi RNA1296 and empty mi RNA-1296 group.CCK-8was used to detect the optimal concentration and action time of drug-containing serum,as well as the effects of Qingre Huashi Decoction and mi RNA1296 on the proliferation of MKN45 cells.The effects of Qingre Huashi Recipe and mi RNA1296 on the apoptosis of MKN45 cells were detected by flow cytometry.The effects of Qingre Huashi Recipe and mi RNA1296 on glucose metabolism of MKN45 cells were detected by glucose and lactic acid kits.Real-time PCR and Western blot were used to detect the effects of Silencing Qingre Huashi Recipe and mi RNA1296 on m TOR and SLC2A3 levels in MKN45 cells.Results:Among the five gastric cancer cells,the expression of mi RNA-1296 in MKN-45 cells was at a moderate level,so MKN-45 was selected as the subsequent overexpression and silencing cells.PCR detection showed that overexpression and silencing of mi RNA-1296 had obvious overexpression and silencing effects compared with the empty mi RNA-1296 group.The results of CCK8 showed that the blank serum with a concentration of 10 %did not affect MKN45 cells after 48 hours of action,and the drug-containing serum with a concentration of 10 % had a significant effect on MKN45 cells after 48 hours of action.Therefore,10 % concentration of blank serum and drug-containing serum were selected,and the action time was 48 hours for subsequent experiments.CCK8 results showed that the drug-containing serum and overexpression of mi RNA-1296 could significantly inhibit the proliferation of MKN45 cells(P<0.01).Silencing mi RNA-1296 could significantly enhance the proliferation of MKN45 cells(P<0.01).Compared with the control group,drug-containing serum and overexpression of mi RNA-1296 could significantly enhance the apoptosis of MKN45 cells(P<0.01).Compared with the control group,drug-containing serum and overexpression of mi RNA-1296 could significantly inhibit the production of glucose and lactic acid in MKN45 cells(P<0.05).Silencing mi RNA-1296 could significantly promote the production of glucose and lactic acid in MKN45 cells(P<0.05).Compared with the control group,the expression of mi RNA-1296 in drug-containing serum and overexpression mi RNA-1296 group was significantly increased(P<0.01),the expression of m TOR and SLC2A3 was significantly decreased(P<0.05),and the expression of mi RNA-1296 in silencing mi RNA-1296 group was significantly decreased(P<0.05).The RNA expression levels of m TOR and SLC2A3 were significantly increased(P<0.01).Compared with the control group,the expression of m TOR and SLC2A3 protein in the drug-containing serum group and the overexpression mi RNA-1296 group was significantly decreased(P<0.01).The expression of m TOR and SLC2A3 protein in the silencing mi RNA-1296 group was significantly increased(P<0.01).Conclusion:mi RNA-1296 may affect the metabolic reprogramming of gastric cancer cells by negatively regulating the m TOR/SLC2A3 pathway,and ultimately interfere with the growth of gastric cancer cells.Qingre Huashi Decoction can up-regulate the expression of mi RNA-1296,inhibit the expression of m TOR and SLC2A3,inhibit the production of glucose and lactic acid,inhibit the proliferation of gastric cancer cells and promote their apoptosis,so as to achieve the synergistic effect of preventing and treating gastric cancer.