Modrowski D, Lomri A, Marie P J
INSERM Unité 349, Cell and Molecular Biology of Bone and Cartilage, Lariboisière Hospital, Paris, France.
J Cell Physiol. 1997 Jan;170(1):35-46. doi: 10.1002/(SICI)1097-4652(199701)170:1<35::AID-JCP5>3.0.CO;2-M.
Granulocyte-macrophage colony-stimulating factor (GM-CSF) is an important modulator of hematopoietic cells. However, the role of GM-CSF in nonhematopoietic cells remains unclear. We have determined whether GM-CSF is an autocrine mitogenic factor for human osteoblastic (hOB) cells. We found by reverse transcriptase-polymerase chain reaction (RT-PCR) that hOB cells express constitutively both GM-CSF and the alpha and beta chains of the GM-CSF receptor (GMR). Immunocytochemistry showed that serum-starved hOB cells express both GM-CSF and GMR alpha chain. Recombinant human (rh) GM-CSF induces a dose-dependent stimulation of hOB cell proliferation, showing that hOB cells have functional GMRs. A specific neutralizing GM-CSF antibody decreased the basal growth rate and suppressed cell proliferation induced by media conditioned by hOB cells, indicating that GM-CSF released by hOB cells is biologically active. Treatment of hOB cells with GM-CSF antisense (AS) oligonucleotide inhibited the endogenous GM-CSF production as shown by ELISA and immunocytochemistry, whereas a random (R) sequence had no effect. AS oligonucleotides markedly inhibited hOB cell growth reversibly, whereas R oligonucleotides had no effect. AS was more effective than the anti-GM-CSF antibody, and the addition of rhGM-CSF did not rescue the inhibitory effect of AS on cell growth. The findings that human osteoblastic cells produce GM-CSF and express functional GMR constitutively and that suppression of endogenous GM-CSF results in inhibition of cell growth demonstrate that GM-CSF is involved as an autocrine growth factor for human osteoblastic cells.
粒细胞-巨噬细胞集落刺激因子（GM-CSF）是造血细胞的重要调节因子。然而，GM-CSF在非造血细胞中的作用仍不清楚。我们已确定GM-CSF是否是人类成骨细胞（hOB）的自分泌促有丝分裂因子。我们通过逆转录聚合酶链反应（RT-PCR）发现，hOB细胞组成性表达GM-CSF以及GM-CSF受体（GMR）的α链和β链。免疫细胞化学显示，血清饥饿的hOB细胞表达GM-CSF和GMRα链。重组人（rh）GM-CSF诱导hOB细胞增殖呈剂量依赖性刺激，表明hOB细胞具有功能性GMR。一种特异性中和GM-CSF抗体降低了基础生长速率，并抑制了hOB细胞条件培养基诱导的细胞增殖，表明hOB细胞释放的GM-CSF具有生物活性。用GM-CSF反义（AS）寡核苷酸处理hOB细胞，如ELISA和免疫细胞化学所示，抑制了内源性GM-CSF的产生，而随机（R）序列则无作用。AS寡核苷酸显著可逆地抑制hOB细胞生长，而R寡核苷酸则无作用。AS比抗GM-CSF抗体更有效，添加rhGM-CSF不能挽救AS对细胞生长的抑制作用。人类成骨细胞产生GM-CSF并组成性表达功能性GMR，以及抑制内源性GM-CSF导致细胞生长受抑制的这些发现表明，GM-CSF作为人类成骨细胞的自分泌生长因子发挥作用。
