New insight into the mechanism of p53 inducing leukemia cell apoptosis.
- Author:
Yuan-zhong CHEN
1
;
Yong WU
;
Min LIANG
;
Nai-nong LI
;
Zhao-xiu ZHANG
;
Lian-huang LU
Author Information
- Publication Type:Journal Article
- MeSH: Apoptosis; drug effects; genetics; DNA, Antisense; pharmacology; DNA-Binding Proteins; Gene Expression Regulation, Neoplastic; HL-60 Cells; Humans; K562 Cells; Leukemia; genetics; pathology; Mutation; Plasmids; genetics; Proto-Oncogene Proteins c-bcl-2; genetics; RNA, Messenger; drug effects; genetics; metabolism; Reverse Transcriptase Polymerase Chain Reaction; Telomerase; genetics; Transfection; methods; Transforming Growth Factor beta; genetics; Transforming Growth Factor beta1; Tumor Necrosis Factor-alpha; genetics; Tumor Suppressor Protein p53; genetics; physiology
- From: Chinese Journal of Hematology 2003;24(12):640-643
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo investigate the expression changes of intrinsic cytokines TGF-beta(1) and TNF-alpha, telomerase activity and bcl-2 during ongoing apoptosis of HL-60 and K562 cells induced by p53.
METHODSpN53cG (Val135), a temperature sensitive p53 mutant, which behaved like wild type p53 (wt-p53) at 32.5 degrees C, were introduced into p53-null HL-60 and K562 cells respectively by lipofectin. In the presence of G418, HL-60-pN53cG and K562 pN53cG clones expressing p53 protein were selected. The ongoing expression of intrinsic cytokines (TGF-beta(1) and TNF-alpha), bcl-2 oncogene and hTERT mRNA during the apoptosis of HL-60 and K562 cells induced by p53 and the effects of exogenous p53 gene, TGF-beta(1) and TNF-alpha antisense PS-ODNS on the apoptosis of HL-60 and K562 cells and the expression of bcl-2 were studied by RT-PCR, quantitative RT-PCR, DNA fragmentation, TdT-mediated dUTP nick end labeling (TUNEL) and flow cytometery. The levels of secreted TGF-beta(1) and telomerase activity were detected by ELISA and PCR-ELISA, respectively.
RESULTS(1) The expressions of intrinsic TGF-beta(1) and TNF-alpha mRNA were up-regulated, while that of bcl-2 and hTERT down-regulated. The levels of TGF-beta(1) in the supernatant of HL-60 and K562 cells were increased, and the level of telomerase activity decreased. (2) Antisense PS-ODNS of TGF-beta(1) and TNF-alpha could obviously inhibit the p53 inducing cell apoptosis, and restore bcl-2 mRNA and protein to pre-treated level.
CONCLUSIONSExogenous p53 induces leukemia cell apoptosis via up-regulating the expression of intrinsic TGF-beta(1) and TNF-alpha and down-regulating the expression of hTERT and bcl-2.
