Objective:The purpose of this study was to observe the anti-cancer activity of Tonglian decoction(TD)on esophageal cancer(EC)cells in vitro,and to elucidate the related molecular mechanisms in the cell cycle and PI3K/Akt signaling pathway.Methods:EC9706 cells were cultured in RPMI 1640 medium supplemented with 10%calf serum at 37C in a 5%CO2 incubator.The cells were treated with rat serum containing TD or the serum of rats administered Xiaoaiping as a positive control drug.Cell proliferation was assessed by methylthiazolyldiphenyl-tetrazolium bromide assays.Cell morphology was observed under a microscope.The cell cycle was examined by flow cytometry.Protein expression in the PI3K/Akt signaling pathway was measured by western blotting.Results:TD mainly inhibited cell proliferation.Concentrations of 50%cell inhibition by rat serum containing TD or Xiaoaiping were 73.6 and 153.8 mL/mL,respectively.TD also influenced cell morphology characterized by small shrunken cells.Cell colonies became small and the cell proliferation rate was slower.In cell cycle analysis,the percentage of cells in S phase was decreased significantly by TD and Xiaoaiping compared with the blank control group(P<.05).Western blotting showed that serum containing TD strongly downregulated EGFR,PI3K,Akt,p-Akt,and mTOR expression compared with the blank control group(P<.05).Conclusion:TD could inhibit EC9706 carcinoma cell proliferation by blocking the cell cycle progression in S phase.The possible mechanism was inhibition of multiple targets in the PI3K/Akt signaling pathway by TD.
Yongsen JiaLijuan QinChunhua JiangQing LinFuling TianHuijuan CaoXin Yan
