Induction of apoptosis in human esophageal cancer cells by sequential transfer of the wild-type p53 and E2F-1 genes: Involvement of p53 accumulation via ARF-mediated MDM2 down-regulation
T. Itoshima et al., Induction of apoptosis in human esophageal cancer cells by sequential transfer of the wild-type p53 and E2F-1 genes: Involvement of p53 accumulation via ARF-mediated MDM2 down-regulation, CLIN CANC R, 6(7), 2000, pp. 2851-2859
Transcriptional factor E2F-1 as well as tumor suppressor p53 have been show
n to cause apoptosis independently in some types of human cancer cells when
overexpressed, Here we report that sequential transfer of the wild-type p5
3 and E2F-1 genes efficiently induces apoptosis in human esophageal cancer
cells and that E2F-1 overexpression directly activates expression of p14 (A
RF), which inhibits MDM2-mediated p53 degradation, resulting in the stabili
zation of p53. Infection of human esophageal cancer cell lines T.Tn and TE8
with adenovirus vector-expressing E2F-1 (Ad-E2F-1) enhanced mRNA and prote
in expression of ARF and decreased MDM2 protein expression. Transfection of
ARF plasmid decreased MDM2 protein expression, which in turn increased p53
protein expression. Infection of T.Tn and TE8 cells first with adenovirus-
expressing wild-type p53 (Ad-p53) and then with Ad-E2F-1 resulted in rapid
induction of apoptosis; in contrast, simultaneous infection with Ad-E2F-1 a
nd Ad-p53 had no significant antitumor effect. As shown by Western blot ana
lysis, infection with suboptimal concentrations of Ad-E2F-1 induced the acc
umulation of exogenous p53 transduced by suboptimal concentrations of Ad-p5
3, Moreover, Ad-E2F-1-mediated ARF expression inhibited the up-regulation o
f MDM2 by overexpressed p53 in TE8 cells. Thus, overexpression of ectopic E
2F-1 protein may stabilize endogenous as well as ectopic p53 protein via th
e E2F-1/ARF/MDM2/p53 regulatory pathway and, in this way, render cells more
sensitive to apoptosis, an outcome that has important implications for the
treatment of human esophageal cancers.