[The effects of WT1 gene down regulation on the sensitivity of K562/A02 cells to adriamycin].
OBJECTIVE
To investigate the effects of WT1 gene down regulation on the sensitivity of K562/A02 cells to adriamycin (ADM).


METHODS
Short hair RNA (shRNA) plasmid vector targeting WT1 gene with enhanced green fluorescent protein (EGFP) was transfected into K562/A02 cells. Transfection efficiency was detected and transfected cells were sorted by flow cytometry. Expression of WT1 gene was detected by real time fluorescence RT-PCR and Western blot. The IC50 of ADM on K562/A02 cells was measured by MTT assay. Intracellular ADM accumulation and ADM-induced apoptosis of K562/A02 cells were detected by flow cytometry.


RESULTS
48 h after transfection,the transfection efficiency was (68.2 +/- 4.5)%. Both the mRNA and protein of WT1 were significantly decreased in K562/A02 cells transfected with WT1 shRNA plasmid than in control. After 24 h ADM induction, the relative reverse rate of sensitivity of K562/A02 cells to ADM was 71.5%. The intracellular accumulation of ADM was greatly increased (P<0.05) and ADM-induced apoptosis rate elevated from 10.4% to 67.3% (P<0.05).


CONCLUSIONS
shRNA targeting WT1 gene can efficiently inhibit the WT1 expression in K562/A02 cells and enhance the drug sensitivity of K562 cells to ADM.
Yang
Tao
T
Institute of Hematology, The Second Hospital of Shanxi Medical University, Taiyuan, China.
Wang
Hong-Wei
HW
Xu
Jing
J
Zhang
Li
L
Zhu
Lei
L
D015536:Down-Regulation; D004317:Doxorubicin / Q000494:pharmacology*; D018432:Drug Resistance, Multiple / Q000235:genetics; D019008:Drug Resistance, Neoplasm / Q000235:genetics*; D015972:Gene Expression Regulation, Neoplastic; D005822:Genetic Vectors; D006801:Humans; D020014:K562 Cells; D012333:RNA, Messenger / Q000235:genetics; D014162:Transfection; D025721:WT1 Proteins / Q000235:genetics* / Q000378:metabolism
D012333:RNA, Messenger; D025721:WT1 Proteins; D004317:Doxorubicin