| Objective:To investigate the expression of AQP9in HepG2human hepatocellular carcinoma and the effect of expression of AQP9on migration of HepG2liver cancer cells, to explore the possible functions and clinical significance of AQP9in the migration of HepG2liver cancer cells.Methods:1.To divide HepG2liver cancer cell lines into three groups.Add oleic acid solution to each group cells according to three concentration gradients(0,250,500μmol/L), which was cultured in a37℃,5%CO2incubator for24hours. Then we extract protein from the cell sample cultured, and test the expression level of AQP9by means of Western blot, the cell migration in Scratch Test.2.To divide HepG2liver cancer cell lines into two groups.The experimental group is dealt with AgN03(3×10^-5) in advance and is cultured in37℃,5%CO2condition.Then we add oleic acid at500μmol/L concentrations into it.The other group,without any intervention, is conducted as control group.After24hours,we extract the sample cells of each group, test the expression level of AQP9by means of Western blot and the cell migration in Scratch Test. Results:1. Western blot showed that relative expression quantity of AQP9in HepG2liver cancer cells with oleic acid at0μmol/,250μmol/L,500μmol/L concentrations induced was (0.679±0.033),(0.804q0.026)and(1.124±0.015) respectively. Relative expression quantity of AQP9in AgN03group and control group was (0.670±0.025),(0.679±0.033).Result indicated that the expression of AQP9in HepG2liver cancer cells was gradually enhanced (P<0.05) with the increase of oleic acid concentration,that is,oleic acid can improve the expression of AQP9in HepG2liver cancer cells,and this effect depended on dose. However,there were no significant differences (P>0.05) on the expression quantity of AQP9in HepG2liver cancer cells between group induced by AgNO3dealt and oleic acid at500μmol/L concentration and control group(with no oleic acid and AgNO3).2.Scratch test showed that the cell migration rate of groups with oleic acid at0μmol/L,250μmol/L,500μmol/L concentrations were (1±0.026),(0.813±0.044) and (0.533±0.079), respectively; While the cell migration rate of AgNO3inhabitor group and control group were (1.008±0.028)、(1±0.026),respectively.Result indicated that migration of HepG2liver cancer cells was gradually enhanced(P<0.05) with the increase of oleic acid concentration.After we added AgNO3to inhabit AQP9,there were no significant differences (P>0.05) on the migration of HepG2liver cells in each group.3. The relationship between expression of AQP9and HepG2liver cancer migration:The increase of expression of AQP9can enhance the migration of HepG2liver cancer cells.But after we use AgNO3to inhabit AQP9, the improvement of HepG2liver cancer cells migration caused by increase of expression of AQP9disappear.Conclusions:1. Oleic acid can improve the expression of AQP9in HepG2liver cancer cells and this improvement depends on dose.2.The increase of expression of AQP9enhances HepG2liver cancer cells migration,which indicates that the expression of AQP9may be related to the migration of HepG2liver cancer. |
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