The Effects Of Polyamines Microenvironment On The Migration Of HCT116 Cells In Vitro

Objective:Polyamines are long-chain aliphatic compounds, including putrescine(Put), spermine(Spm) and spermidine(Spd). Polyamines are widely found in nature, which is necessary for many vital organs. Physiological polyamine content in human tissues and body fluids is very low. But the content of many polyamines content in tumor tissue is more than in normal tissue which is related to tumor genesis, development and metastasis.The aim of this experiment was to evaluate the effects of different polyamine microenvironmen(high, normal and low) on cell growth and metastasis of colon carcinoma cell and its molecular mechanisms in vitro.Methods:Akt1 gene was transfected into HCT116 cells, named HCT116-Akt1 cells, by gene transfection technique. The different exogenous polyamines, including high concentrations of polyamines(putrescine 0.8 nmol / mL, spermidine 10.0 nmol / mL, spermine 5.0 nmol / mL); normal(middle) concentration of polyamines(putrescine 0.08 nmol / mL, spermidine 1.0 nmol / mL, spermine 0.5 nmol / mL); low concentration of polyamines(putrescine 0.008 nmol / mL, spermidine 0.1 nmol / mL, spermine 0.05 nmol / mL) were added to HCT116 cells and HCT116-Akt1 cells. HCT116 cells and HCT116-Akt1 cells were cultured for 15 or 30 days in the exogenous polyamines. Colony-forming ability was detected to evaluat the cells proliferation; Transwell assay and wound healing capability were examined to assess the cells adhesion, migration and invasion. Western blot was used to investigate the protein expression including AKT1, AKT, ODC, MMP2, Integrinα6 and SSAT.Results:Polyamine microenvironment influenced the ability of proliferation and migration on HCT16 cells and HCT116-Akt1 cells in vitro. Furthermore, higher polyamine microenvironment significantly facilitated the proliferation and migration of HCT116-Akt1 cells than HCT116 cells in a dose- and time-dependent manner. Western blot demonstrated that polyamine affected the protein expression of Akt1, ornithine decarboxylase(ODC), spermidine and spermine acetyltransferase(SSAT), Integrin α6.Conclusion:Polyamine microenvironment affected facilitate the proliferation and metastasis of HCT116 cells and HCT116-Akt1 cells in vitro. Furthermore, the proliferation and metastasis of HCT116-Akt1 cells were significantly enhanced compared with HCT116 cells, suggesting that Akt1 plays a vital role in polyamine-mediated colon carcinoma metastasis.