Effect of Huqi San Contained Serum on Glycolysis of Esophageal Cancer Cells Ec9706 and Regulation of AMPK/FOXO3a Pathway
Objective To study the effects of Huqi San contained serum on glycolysis of esophageal cancer cells Ec9706 and the regulation of AMPK/FOXO3a pathway.Methods Ec9706 cells were collected and inoculated in cell culture dishes.Blank control group,Huqi San low-dose,medium-dose and high-dose groups and cisplatin groups were set up,respectively.Each dose group of Huqi San was added Huqi San contained serum,cisplatin group was added 2 μg·mL-1 cisplatin,and blank control group was added fresh DMEM culture medium.The proliferation rate of Ec9706 cells was determined by MTT assay,the glucose intake,lactic acid production,HK and LDH activities of Ec9706 cells were determined by ELISA assay,the 24 h mobility of Ec9706 cells was determined by scratch assay,and the invasion rate of Ec9706 cells was determined by Transwell assay.The apoptosis rate of Ec9706 cells was determined by flow cytometry,and the mRNA levels of FOXO3a,Bax,Bcl-2 and LDHA in Ec9706 cells were determined by RT-qPCR.The protein levels of p-AMPK,FOXO3a,Bax,Bcl-2 and LDHA in Ec9706 cells were determined by Western blot.Results Compared with blank control group,the proliferation rate,glucose intake,lactate production,HK activity and LDH activity of Ec9706 cells at 24 h,48 h and 72 h in Huqi San different doses groups and cisplatin group,the migration rate,invasion rate,Bcl-2 and LDHA mRNA and protein levels,p-AMPK/AMPK levels of Ec9706 cells at 24 h were significantly decreased(P<0.05).Apoptosis rate of Ec9706 cells,the mRNA and protein levels of FOXO3a and Bax were significantly increased(P<0.05),and with the increase of the dose of Huqi San,the changes of various indexes of Ec9706 cells were better.Conclusion Huqi San can significantly inhibit the glycolysis of esophageal cancer Ec9706 cells,and inhibit Ec9706 cells proliferation,and promote the apoptosis of Ec9706 cells.The mechanism may be related to the regulation of AMPK/FOXO3a pathway.
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