[Effect of the new human transcription factor hBKLF on the proliferation, differentiation of K562 cell line and hemoglobin synthesis].
The human basic Krüppel-like factor (hBKLF) is a newly cloned human transcription factor from the cDNA library of fetal liver. It belongs to the Krüppel-like transcription factor family. Previous expression study showed that it is a hematopoietic related factor. This study was aimed to investigate the effect of hBKLF on cell proliferation, differentiation and hemoglobin synthesis by using K562 cell line as model. The sense and antisense expression plasmids of hBKLF were constructed, and transfected into K562 cells by lipofectamine. After G418 selection for 4 weeks, the cell line with stable expression of the gene was obtained. Then the hBKLF expression level, proliferation ability, colony formation and hemoglobin production were detected by RT-PCR and Western blot, MTT method, methyl cellulose semisolid culture method and benzidine test respectively. The morphologic change of cell was observed with inverted microscope. The results showed that the sense plasmid could increase hBKLF level and antisense plasmid could decrease hBKLF expression. When hBKLF level was down-regulated, K562 cells could proliferate more quickly and synthesize more hemoglobin. But there were no differences in colony formation ability and no apparent morphologic change. It is concluded that hBKLF can inhibit hematopoietic cell proliferation and hemoglobin synthesis. It is suggested that hBKLF plays an important role in the proliferation and differentiation of hematopoietic cells.
Wang
Mang-Ju
MJ
Department of Hematology, Peking University First Hospital, Beijing 100034, China.
Ma
Xiao-Yun
XY
Shi
Yong-Jin
YJ
Wu
Shu-Lan
SL
He
Fu-Chu
FC
D000818:Animals; D019556:COS Cells; D002454:Cell Differentiation / Q000502:physiology*; D049109:Cell Proliferation / Q000187:drug effects*; D002471:Cell Transformation, Neoplastic / Q000187:drug effects; D002522:Chlorocebus aethiops; D006454:Hemoglobins / Q000096:biosynthesis*; D006801:Humans; D020014:K562 Cells; D051741:Kruppel-Like Transcription Factors / Q000096:biosynthesis / Q000235:genetics* / Q000494:pharmacology; D014157:Transcription Factors / Q000096:biosynthesis / Q000235:genetics; D014162:Transfection
D006454:Hemoglobins; D051741:Kruppel-Like Transcription Factors; D014157:Transcription Factors