Kamano H, Ohnishi H, Tanaka T, Ikeda K, Okabe A, Irino S
First Department of Internal Medicine, Kagawa Medical School, Japan.
Leuk Res. 1990;14(10):831-9. doi: 10.1016/0145-2126(90)90171-5.
Recombinant plasmids containing v-myb' (803 bp fragment of the 3' end of v-myb) were constructed to induce sense or antisense v-myb' RNA expression with dexamethasone in human cells. These plasmids were used as a tool for the investigation of the role of c-myb gene in human leukemia cell proliferation and differentiation. They were transfected by electroporation into the K562 human leukemia cell line derived from a patient with chronic myelogenous leukemia in blastic crisis. After induction of transcription by dexamethasone, the plasmid with antisense v-myb' repressed the expression of p75c-myb from the endogenous c-myb gene of K562 cells. It also reduced the proliferation rate of K562 cells to 50% of the control level, and induced these K562 cells to express the myelomonocytic differentiation cell surface marker CD13 and increased NBT reducing activity. The plasmid with sense v-myb' did not have an effect on p75c-myb expression, the proliferation of K562 cells or the expression of myelomonocytic differentiation phenotypes. These observations suggest that antisense v-myb' RNA represses p75c-myb expression and that a decrease of p75c-myb suppresses K562 cell proliferation and induces its differentiation towards the myelomonocytic lineage.
构建了含有v-myb'(v-myb 3'端803 bp片段)的重组质粒,以用地塞米松在人细胞中诱导正义或反义v-myb' RNA表达。这些质粒用作研究c-myb基因在人白血病细胞增殖和分化中作用的工具。通过电穿孔将它们转染到源自一名处于急变期的慢性粒细胞白血病患者的K562人白血病细胞系中。用地塞米松诱导转录后,带有反义v-myb'的质粒抑制了K562细胞内源性c-myb基因的p75c-myb表达。它还将K562细胞的增殖率降低至对照水平的50%,并诱导这些K562细胞表达髓单核细胞分化细胞表面标志物CD13,同时提高了NBT还原活性。带有正义v-myb'的质粒对p75c-myb表达、K562细胞增殖或髓单核细胞分化表型的表达没有影响。这些观察结果表明,反义v-myb' RNA抑制p75c-myb表达,并且p75c-myb的减少抑制K562细胞增殖并诱导其向髓单核细胞系分化。