[Expression of WT1 gene and its isomer ratio changes during phorbol ester induced differentiation of K562 cell line].
OBJECTIVE
To explore the changes in expression of WT1 gene and ration of its isomers during phorbol ester (TPA) induced differentiation of leukemia cell line K562 by fluorescence quantitative RT-PCR and analysis the relationship between different isomers and hematogenic cell differentiation.


METHODS
The degree of cellular maturation were verified by NBT reduction test and immunophenotyping. Expression of WT1 gene was determined by fluorescence quantitative RT-PCR during differentiation of K562 cell line. The relative ratio of the four splicing variants WT1 ( + / + ), WT1 ( + / - ), WT1 ( - / + ), WT1 ( - / - ) were calculated.


RESULTS
During the differentiation of K562 cell, the NBT reduction rate and the CD9 positive rate both increased significantly (P < 0. 05). The expression of WT1 gene decreased immediately to (1.67 +/- 0.45) x 10(-3) from (4.67 +/- 1.11) x 10(-3), and then increased again to (4.64 +/- 1.53) x 10(-3) at 96 hours. The ratio of WT1 ( + / + ) was decreased gradually, from 0 hour (39.65 +/- 19.46)% to 96 hour (15.25 +/- 7.27)%. While the ratio of WT1( - / - ) was increased, from 0 hour (15.38 +/- 11.34)%, to 96 hour (37.60 +/- 11.90)%. The other two isomers ratios did not change significantly.


CONCLUSIONS
During the TPA induced differentiation of K562 cell, there are two high expression levels of WT1 gene. Before differentiation, the majority is WT1 ( + / + ), and after differentiation, is WT1 ( - / - ). It indicates that WT1 gene may activate or inhibit cell differentiation by regulating the ratio of its four splicing variants.
Li
Xiao-hong
XH
Institute of Hematology, The Second Hospital of Shanxi Medical University, Taiyuan 030001, China.
Wang
Hong-wei
HW
Li
Jian-lan
JL
Zhu
Lei
L
Fan
Wei-ping
WP
Tian
Cai-xia
CX
Xu
Jing
J
Xu
Yong-qun
YQ
D002454:Cell Differentiation / Q000187:drug effects* / Q000235:genetics; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D020014:K562 Cells; D010703:Phorbol Esters / Q000494:pharmacology*; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism; D025721:WT1 Proteins / Q000235:genetics / Q000378:metabolism*
D010703:Phorbol Esters; D020033:Protein Isoforms; D025721:WT1 Proteins