Effects of 3-bromopyruvate on the proliferation,migration,and invasion of Hep2 laryngeal carcinoma cells via the PI3K/Akt/mTOR signaling pathway
Objective:3-Bromopyruvate(3-BrPA)is a novel potential anticancer agent that inhibits glycolytic and has gained considerable attention in cancer treatment.However,its effects in laryngeal cancer remain unclear.This study investigates the impact of 3-BrPA on the proliferation,migration,and invasion of human Hep2 laryngeal cancer cells and explores its potential mechanisms via the phosphoinositide 3-kinase(PI3K)/protein kinase(Akt)/mammalian rapacity target protein(mTOR)signaling pathway.Methods:Human Hep2 laryngeal cancer cells were cultured in vitro and divided into a control group and 3-BrPA(20,40,60,80,100,120 μmol/L)treatment groups for 24,48,and 72 hours to observe the effects on cell proliferation.Optimal concentrations and exposure times were determined for further experiments.Scratch and Transwell assays were used to assess cell migration and invasion.The STRING database was used to analyze correlations between hexokinase-II(HK-2)and PI3K,Akt,and mTOR proteins.Real-time RT-PCR and Western blotting analyses were performed to measure mRNA and protein expression of HK-2,PI3K,Akt,and mTOR.Results:Cell counting-kit 8(CCK-8)assays indicated a significant decrease in proliferation of Hep2 cells exposed to 40-120 μmol/L 3-BrPA at 24,48,and 72 hours compared to the control(all P<0.05).Concentrations of 95,115,and 135 μmol/L were selected as low,medium,and high for subsequent 24-hour treatment experiments.CCK-8 results revealed that as 3-BrPA concentration increased,Hep2 cell proliferation,migration rate,and invasion number significantly decreased compared to the control group(all P<0.01).STRING analysis indicated a strong correlation between HK-2 and Akt/mTOR proteins in Hep2 cells.Real-time RT-PCR and Western blotting results demonstrated that HK-2,PI3K,Akt,and mTOR mRNA and protein expression significantly decreased in the low,medium,and high 3-BrPA treatment groups compared to the control(all P<0.05).Conclusion:3-BrPA inhibits the proliferation,migration,and invasion of Hep2 laryngeal cancer cells,potentially through down-regulation of the PI3K/Akt/mTOR signaling pathway.