Figure 10.

E-cadherin expression and promoter activity in iron-loaded human hepatic cell cultures. A: E-cadherin protein expression was decreased in Huh7 cells treated with FeNTA. The Huh7 human hepatoma cell line was treated with 100 μmol/L of FeNTA for 72 or 96 hours. Twenty-four hours after plating, Huh7 cells were treated, in the absence of fetal bovine serum, with control medium or medium supplemented with 100 μmol/L of FeNTA. Cultures were either not refed (72 hours or 96 hours) or refed every 24 hours (72 hours × 3 or 96 hours × 4) with the appropriate medium. After 72 hours or 96 hours, cultures were harvested for protein and analyzed for E-cadherin or ferritin protein expression. B: E-cadherin promoter activity was decreased in FeNTA-treated human hepatic cells. Huh7 cells were transfected with either CMV-renilla plasmid alone (control), CMV-renilla and pGL3 plasmids (pGL3), or CMV-renilla and pGL3/PECAD (pGL3/PECAD) plasmids in serum-free medium in the absence or presence of 100 μmol/L of FeNTA (+FeNTA). At 24 hours after transfection, cultures were harvested and analyzed for relative (firefly/renilla) luciferase activity (RLU).