Mechanism of Compound Fuling Granule Regulating Glucose Metabolism Reprogramming and Inhibiting Ovarian Cancer Me-tastasis through DRP1 / 浙江中医药大学学报

ABSTRACT

[Objective]To investigate the mechanism of Compound Fuling Granule(CFG)in inhibiting the glucose metabolism and metastasis of ovarian cancer cells.[Methods]Ovarian cancer cell HEY-T30 were cultured in vitro and incubated with different concentration(0.25,0.5,1,1.5,2 mg·mL-1)of CFG for 24 h.Cell counting kit-8(CCK-8)assay was performed to detect the effect of CFG on the cell proliferation,Transwell assay was used to investigate cell migration ability,lactic acid detection kit and glucose detection kit were used to detect lactic acid production and glucose consumption,the expressions of dynamin-related protein 1(DRP1),some key glycolysis-related proteins and metastasis-related proteins were detected by Western blot,Mito-Tracker Red was used to label mitochondria to observe mitochondria morphology.Lentivirus transfection technique was used to achieve the stable DRP1 knockdown HEY-T30 cells.Real-time quantitative polymerase chain reaction(Real-time qPCR)was used to detect the expression of DRP1 mRNA,the effect of CFG on lactic acid production and glucose consumption of HEY-T30 after DRP1 knockdown was detected by lactic acid detection kit and glucose detection kit,Transwell assay was used to investigate the migration ability of HEY-T30 with DRP1 knockdown after treated with CFG,the effect of CFG on the expression of glycolysis-related proteins and metastasis-related proteins in HEY-T30 with DRP1 knockdown was detected by Western blot.[Results]Compared with control group,the cell survival rate in 0.5,1,1.5,2 mg·mL-1 CFG groups reduced significantly(P＜0.01).The average length of mitochondria in 0.5,1 mg·mL-1 group was markedly increased(P＜0.01),the protein expression of DRP1 was significantly reduced(P＜0.05,P＜0.01),and the lactate production and glucose consumption in 0.5 and 1 mg·mL-1 groups were significantly decrease(P＜0.01).The number of migration cell was significantly reduced in 0.25,0.5 and 1 mg·mL-1 concentration groups(P＜0.05,P＜0.01).After knockdown of DRP1,the glycolysis level and migration of HEY-T30 were decreased(P＜0.05,P＜0.01),and the expressions of glycolysis-related proteins and metastasis-related proteins were decreased(P＜0.05,P＜0.01).The inhibitory effect of CFG on glycolysis and metastasis of ovarian cancer cells was also weakened.[Conclusion]By targeting DRP1 to regulate glucose metabolism reprogramming,CFG could inhibit the metastasis of ovarian cancer.