The parental cell line 253T was established from a 2acetylaminofluorene induced rat hepatocellular carcinoma, as described previously [35] . The anchorage dependent 253T-NT cell line was isolated from 253T by limiting dilution cloning. 253T and 253-NT cells were grown in Waymouth medium (Sigma, St. Louis, MO, USA) supplemented with 15% FBS, 1% glutamine (Invitrogen), 0.1% Gentamycin (Invitrogen), and 0.2% Normocin (Invivogen, San Diego, CA, USA). For cell proliferation assays, 1.5610 4 cells were plated in a 24-well plate. At 24, 48, 72, and 96 hours after plating, cells were trypsinized (Invitrogen), stained with trypan blue and counted using a hemocytometer.