论文部分内容阅读
HEL cells, a human erythroleukemia cell line, mainly express the fetal (r)globin gene and trace amount of the embryonic (E)globin gene, but not adult (B) globin gene. Here we show that hydroxyurea (HU) can induce HEL cells to express adult (B) globin gene and lead these cells to terminal differentiation. Results showed in Gel mobility shift assays that GATA factors could specifically bind to the regulatory elements of human B- globin gene, including the proximal regulatory element (the B- promoter) and the distal regulatory elements (the DNase I hypersensitive sites in the LCR, HS2-HS4 core sequences). However, the DNA binding patterns of GATA factors were quite different between HU-induced and uninduced HEL cells. Western-blot analysis of nuclear extracts from both the uninduced and HU- induced HEL cells revealed that the level of GATA-2 transcription factor decreased, whereas the level of GATA-1 transcription factor increased following the time of hydroxyurea induction. Furthermore, using RT-PCR analysis the expression of human B-globin gene in HU-induced HEL cells could be blocked again when HEL cells were incubated in the presence of antisense oligonucleotides for hGATA-1, suggesting that the upregulation of hGATA-1 transcription factor might be critical for the expression of human β- globin gene in HU-induced HEL cells.
HEL cells, a human erythroleukemia cell line, mainly express the fetal (r) globin gene and trace amount of the embryonic (E) globin gene, but not adult (B) globin gene. Here we show that hydroxyurea Cells in express adult (B) globin gene and lead these cells to terminal differentiation. Results in Gel mobility shift assays that GATA factors could specifically bind to the regulatory elements of human B-globin gene, including the proximal regulatory element (the B- promoter and the distal regulatory elements (the DNase I hypersensitive sites in the LCR, HS2-HS4 core sequences). However, the DNA binding patterns of quite different between HU-induced and uninduced HEL cells. Western-blot analysis of nuclear extracts from both of uninduced and HU-induced HEL cells revealed that the level of GATA-2 transcription factor decreased, whereas the level of GATA-1 transcription factor increased by the time of hydroxyurea induction. using RT-PCR analysis the expression of human B-globin gene in HU-induced HEL cells could be blocked again when HEL cells were incubated in the presence of antisense oligonucleotides for hGATA-1, suggesting that the upregulation of hGATA-1 transcription factor might be critical for the expression of human β-globin gene in HU-induced HEL cells.