E1AF Induces Both Matrix Metalloprotease Transcription and Cell Cycle Arrest that Occurs in the Stage of Cancer Cell Invasion

Abstract

and to elucidate the relationship between tumor invasive ability and cell cycle modulation caused by E1AF. Materials and Methods Reconstruction of the expression vectorThe E1AF expression vector with a metallothionein promoter was reconstructed as follows.The full-length of E1AF cDNA was cut out by EcoR1, then BamH1 linker ligation into the pSVneoHMTTer metallothionein inducible expression vector was carried out. The vector (phMT II E1AF) contains a neomycin-resistant selective gene sequence, and ligated E1AF was transcribed and translated by adding ZnCl 2 to the medium using the hMT II-metallothionein promoter. Cells and cell cultureThe human oral squamous cell carcinoma-derived cell line Ca9.22 was used for transfection of the phMT II E1AF vector. Cells were maintained in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum (Filtron Pty Ltd, Brooklin, Australia) and antibiotics.