The influence of different WT1 gene isoforms expression pattern on the differentiation of leukemia cell line NB4.
- Author:
Hui-ling SHEN
1
;
Zi-xing CHEN
;
Wei WANG
;
Jian-nong CEN
;
Shao-yan HU
;
Ye ZHAO
Author Information
- Publication Type:Journal Article
- MeSH: Cell Cycle; Cell Differentiation; genetics; Cell Line, Tumor; Cyclin-Dependent Kinase Inhibitor p21; metabolism; Gene Expression Regulation, Neoplastic; Genetic Vectors; Humans; Leukemia, Promyelocytic, Acute; genetics; metabolism; pathology; Proto-Oncogene Proteins c-myc; metabolism; RNA, Messenger; genetics; Transfection; WT1 Proteins; genetics; metabolism
- From: Chinese Journal of Hematology 2005;26(9):543-547
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo study the potential effects of exogenous WT1 gene isoform on the differentiation of leukemia cell line NB4 and its possible molecular mechanisms.
METHODSThe recombinant eukaryotic expression vector (pCB6 + /WTA) containing full-length human WT1 isoform (WTA: -17AA/ -KTS) cDNA and the blank pCB6 + vector were transfected into leukemia cell line NB4 by electroporation. The WTA mRNA and protein in cells were detected by RT-PCR and Western blot. Cell morphology, NBT reduction and CD11b antigen expression in NB4 cells were assayed to evaluate cell differentiation. Expression of PML/RARalpha, p21 and c-myc genes was determined by semi-quantitative RT-PCR after transfection.
RESULTSCompared with NB4/WTA cells, NB4 and NB4/CMV (NB4 cells transfected with pCB6 + vector) cells had higher morphological differentiation rates and higher CD11b expression levels after exposure to ATRA for 48 hours. The percentage of NBT reduction in NB4/WTA cells was lower than that in control groups. The difference in NBT reduction rate between NB4/WTA and control cells was gradually increased after treated with ATRA for three days. The expression levels of PML/RARalpha, p21 and c-myc genes in NB4/WTA cells were notably increased.
CONCLUSIONOverexpression of exogenous WTA gene could partially inhibit the differentiation of NB4 cells by up-regulating the expression of PML/RARalpha, p21 and c-myc genes.
