Holt J T, Redner R L, Nienhuis A W
Clinical Hematology Branch, National Heart, Lung, and Blood Institute, Bethesda, Maryland 20892.
Mol Cell Biol. 1988 Feb;8(2):963-73. doi: 10.1128/mcb.8.2.963-973.1988.
To study the role of a nuclear proto-oncogene in the regulation of cell growth and differentiation, we inhibited HL-60 c-myc expression with a complementary antisense oligomer. This oligomer was stable in culture and entered cells, forming an intracellular duplex. Incubation of cells with the anti-myc oligomer decreased the steady-state levels of c-myc protein by 50 to 80%, whereas a control oligomer did not significantly affect the c-myc protein concentration. Direct inhibition of c-myc expression with the anti-myc oligomer was associated with a decreased cell growth rate and an induction of myeloid differentiation. Related antisense oligomers with 2- to 12-base-pair mismatches with c-myc mRNA did not influence HL-60 cells. Thus, the effects of the antisense oligomer exhibited sequence specificity, and furthermore, these effects could be reversed by hybridization competition with another complementary oligomer. Antisense inhibition of a nuclear proto-oncogene apparently bypasses cell surface events in affecting cell proliferation and differentiation.
为了研究一种核原癌基因在细胞生长和分化调控中的作用,我们用一种互补反义寡聚物抑制HL - 60细胞的c - myc表达。这种寡聚物在培养中稳定且能进入细胞,形成细胞内双链体。用抗myc寡聚物孵育细胞可使c - myc蛋白的稳态水平降低50%至80%,而对照寡聚物对c - myc蛋白浓度无显著影响。用抗myc寡聚物直接抑制c - myc表达与细胞生长速率降低及髓系分化诱导有关。与c - myc mRNA有2至12个碱基对错配的相关反义寡聚物对HL - 60细胞无影响。因此,反义寡聚物的作用表现出序列特异性,而且,这些作用可通过与另一种互补寡聚物的杂交竞争而逆转。核原癌基因的反义抑制在影响细胞增殖和分化时显然绕过了细胞表面事件。