Antisense SCL suppresses self‐renewal and enhances spontaneous erythroid differentiation of the human leukaemic cell line K562.
Antisense SCL suppresses self‐renewal and enhances spontaneous erythroid differentiation of the human leukaemic cell line K562.
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DOI：
10.1002/j.1460-2075.1991.tb04993.x
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发表时间：
1991-12
期刊：
影响因子：
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通讯作者：
A. Green;E. DeLuca;C. Begley
中科院分区：
文献类型：
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作者：
A. Green;E. DeLuca;C. Begley
SCL基因编码螺旋-环-螺旋转录因子家族的成员，这些转录因子参与了分化和发育的调节。虽然SCL mRNA在正常胸腺细胞或外周T淋巴细胞中检测不到，但转录激活发生在T细胞肿瘤中。SCL正常功能的线索来自于红系细胞中高水平的SCL mRNA的证明。为了阐明SCL在红系中的功能，将反义SCL构建体引入人红白血病细胞系K562中。用含有反义SCL的载体电穿孔的细胞比仅接受载体的对照细胞生长更慢。通过显示反义SCL不影响Raji细胞（一种不表达内源性SCL mRNA的B细胞系）的生长，排除了非特异性毒性。抑制K562的生长伴随着增加自发性红细胞分化，通过联苯胺染色测量。与对照细胞相比，含反义SCL的K562细胞在琼脂中产生较小的集落，并表现出降低的集落形成性。此外，K562集落再克隆的实验表明，反义SCL深刻地抑制了K562细胞的自我更新。这些数据提供了SCL促进红系细胞系自我更新的第一个证据，并提高了SCL可能用于调节正常红系细胞增殖的可能性。
The SCL gene encodes a member of the helix‐loop‐helix family of transcription factors that have been implicated in regulation of differentiation and development. Although SCL mRNA is not detectable in normal thymocytes or peripheral T‐lymphocytes, transcriptional activation occurs in T‐cell tumours. A clue to the normal function of SCL has come from demonstration of high levels of SCL mRNA in erythroid cells. To illuminate the function of SCL in the erythroid lineage, an antisense SCL construct was introduced into the human erythroleukaemia cell line, K562. Cells electroporated with a vector containing antisense SCL grew more slowly than control cells which had received vector alone. Non‐specific toxicity was excluded by showing that antisense SCL did not influence growth of Raji cells, a B‐cell line that does not express endogenous SCL mRNA. Suppression of K562 growth was accompanied by increased spontaneous erythroid differentiation as measured by benzidine staining. K562 cells containing antisense SCL produced smaller colonies in agar and exhibited reduced clonogenicity compared with control cells. In addition, experiments in which K562 colonies were recloned showed that antisense SCL profoundly suppressed self‐renewal of K562 cells. These data provide the first evidence that SCL promotes self‐renewal in an erythroid cell line and raise the possibility that SCL may function to regulate proliferation of normal erythroid cells.