[Biological characteristics of Wt1 gene in relation to Ph(+) leukemia cell line K562].
Wt1 is a dual-function gene involved in hematopoiesis, leukemogenesis and prognosis for leukemia. This gene is highly expressed in acute myeloid leukemia (AML) and the progression of chronic myelogenous leukemia (CML). It was reported elsewhere that high level of wt1 expression indicated worse prognosis for leukemia. Wt1 gene functions are different due to its subcellular localization. This study was aimed to investigate the expression and localization of wt1 mRNA and WT1 protein, and explore the effects of wt1 inhibitor, curcumin, on K562 cell proliferation, cell cycle and its possible mechanisms. MTT method was used to detect cell proliferation; flow cytometry was used to analyze the alteration of cell cycle, and the immunofluorescence and Western blot technology were performed to observe the subcellular localization of WT1 protein. The transcripts of wt1 and bcr/abl p210 was analyzed by real-time PCR. The results showed that wt1 mRNA and its protein were both highly expressed in K562 cells. The curcumin and imatinib (Glevec) both inhibit the cell proliferation resulting in the G(2)/M and G(0)/G(1) phase arrest respectively. Meanwhile, the transcripts of wt1 and bcr/ablp210 genes decreased greatly after being treated with the two inhibitors above. It is concluded that the alteration of wt1 gene affects the biological characteristics of Ph(+) K562 cells, such as cell proliferation, cell cycle and so on. Gene wt1 is expected to be further studied as a new therapy target in Ph(+) leukemias.
Li
Yan
Y
State Key Laboratory of Experimental Hematology, Institute of Hematology & Blood Disease Hospital, Chinese Academy of Medical Sciences & Peking Union of Medical College, Tianjin 300020, China.
Li
Xiao-Yan
XY
Wang
Lin
L
Tian
Zheng
Z
Rao
Qing
Q
Jia
Hai-Rong
HR
Wang
Hui-Jun
HJ
Wang
Min
M
Mi
Ying-Chang
YC
D002453:Cell Cycle; D049109:Cell Proliferation; D003474:Curcumin / Q000494:pharmacology; D006801:Humans; D020014:K562 Cells; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D012333:RNA, Messenger / Q000235:genetics; D025721:WT1 Proteins / Q000235:genetics*
D012333:RNA, Messenger; D025721:WT1 Proteins; D003474:Curcumin