[Establishment and characterization of leukemic cell line U937 stably expressing exogenous RbAp46].
To establish leukemic cell lines stably transfected by RbAp46 gene, electroporation was performed after optimizing the transfection condition for suspended cells. Under conditions of low voltage and high capacitance, RbAp46 was transfected into U937 by electroporation. Individual clones selected with G418 for 3 weeks were isolated. The integration and the protein levels of the exogenous RbAp46 in transfectants were determined by PCR and Western blot analysis, respectively. The subclone expressing high level of RbAp46 was then established. Viability of transfected cells was assayed by trypan blue exclusion. Cell number was counted daily to determine the growth rate. The results showed that growth rate of U937 cell lines expressing exogenous RbAp46 was about 50% lower than that in control. It is concluded that leukemic cell lines stably expressing exogenous RbAp46 were established and overexpression of RbAp46 inhibits the growth of U937 leukemic cells.
Duan
Wei-Ming
WM
Jiansu Institute of Hematology, The First Affiliated Hospital of Suzhou University, Suzhou 215006, China.
Chen
Zi-Xing
ZX
Wang
Wei
W
Fu
Jian-Xin
JX
Bai
Xia
X
Zhao
Xiao-Juan
XJ
Yao
Li
L
D015153:Blotting, Western; D002352:Carrier Proteins / Q000235:genetics*; D049109:Cell Proliferation; D018274:Electroporation; D006801:Humans; D009687:Nuclear Proteins / Q000235:genetics*; D056524:Retinoblastoma-Binding Protein 7; D014162:Transfection; D020298:U937 Cells; D025721:WT1 Proteins / Q000235:genetics
D002352:Carrier Proteins; D009687:Nuclear Proteins; C490746:RBBP7 protein, human; D056524:Retinoblastoma-Binding Protein 7; D025721:WT1 Proteins