Influence Of AQP9 Expression Level On Anti-Hepatoma Effects Of Arsenic Trioxide

OBJECTIVE:To investigate the effect of AQP9 expression level on the arsenic trioxide(As2O3)-induced apoptosis and biological behaviors of hepatocellular carcinoma cell line HepG2 by changing the aquaglyceroporin 9(AQP9) expression with recombinant plasmid transfection.METHODS:(1)The effects of As2O3 with different concentration on the cell proliferation was measured by methylthiazolyl tetrazolium (MTT). IC50 was calculated. (2)The recombinant plasmid pEGFP-N1 AQP9 and pshRNA-AQP9a were transfected into HepG2 cells.The expression of AQP9 mRNA and protein was detected by fluorescent microscopy,RT-PCR and Western blowing,respectively.(3)Added As2O3 to transfected and untreated HepG2 cells.The proliferation was tested by MTT. The cell cycle and apoptosis rate was measured by flow cytometry. The ability of the cell invasion and migration was measured by transwell chamber experiment. The activity of Caspase-3 was detected by Microplate reader.RESULTS:(1)The expression of green fluorescent protein was observed by fluorescence microscope. Recombinant plasmid could remarkably enhance and decrease the expression of AQP9. (2)The cell proliferation invasion and migration of hepatocellular carcinoma cell lines transfected pEGFP-N1-AQP9 were attenuated significantly compared with group treated with drug alone. The augmented population of apoptotic cells、increased caspase-3 activity and the highest percentage of cells stagnating at G0/G1 phase were observed in hepatocellular carcinoma cell lines transfected pEGFP-N1-AQP9 compared with group treated with drug alone.(3)The proliferation,invasion and migration of Hepatocellular carcinoma cell lines transfected pshRNA-AQP9a were increased compared with group treated with drug alone. The reduced population of apoptotic cells,attenuated caspase-3 activity and decreased percentage of cells stagnating at G0/G1 phase were found in carcinoma cell lines transfected pshRNA-AQP9 compared with group treated with drug alone.CONCLUSIONS:Proliferation inhibition and apoptosis rate of hepatocellular carcinoma cell lines transfected pEGFP-N1-AQP9 were enhanced in pEGFP-N1-AQP9-transfected group compared with group treated with As2O3 alone. Increasing the level of AQP9 expression can improve the sensitivity to As2O3 in hepatocellular carcinoma cell lines HepG2 and further affect the clinical efficacy of As2O3 in Hepatocellular carcinoma treatment.