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急性髓性白血病中白细胞介素-6(IL-6)受体基因的表达及白血病细胞对外源性IL-6的反应。细胞系细胞与相应原代细胞的比较研究。

Expression of interleukin-6 (IL-6) receptor gene in acute myeloblastic leukemia and response of leukemic cells to exogenous IL-6. A comparative study between cell line cells and corresponding native cells.

作者信息

Säily M, Koistinen P, Savolainen M, Rantala M, Savolainen E R

机构信息

Department of Clinical Chemistry, University of Oulu, Finland.

出版信息

Growth Factors. 1998;15(4):243-57. doi: 10.3109/08977199809017481.

Abstract

As interleukin (IL-6) ahs been reported to have diverse effects on acute myeloblastic leukemia (AML) blast cell growth, we investigated whether the level of IL-6 receptor (IL-6R) expression by blast cells is associated with their susceptibility to proliferate in response to exogenous IL-6. For absolute quantification of IL-6R transcript numbers, we established a quantitative IL-6R reverse transcription polymerase chain reaction (RT-PCR) method with an internal RNA standard. In the present work, two types of AML blast cells were investigated, namely autonomously growing cell line cells (n = 8) and non cultured native blast cells (n = 20), including those from which the cell lines originate. The native blast cells expressed an average of 2.8 x 10(7) +/- 1.9 x 10(7) IL-6R transcripts in one microgram of total cellular RNA, whereas the expression by the cell line cells was significantly more abundant, the value being 8.3 x 10(7) +/- 2.8 X 10(7) (P < 0.001). The proliferation responses were evaluated by exposing the cells to IL-6 (1000 U/ml) in a clonogenic cell culture assay and, in the case of the cell line cells, in a long-term suspension culture assay as well. None of the autonomously growing cell lines responded to exogenous IL-6, whereas the native blast cell showed either stimulatory, inhibitory or neutral responses. Thus, the IL-6R expression level did not predict whether the cells proliferated in response to exogenous IL-6, which shows that IL-6R quantification cannot be used as a screening test prior to possibly applying this cytokine to clinical use in AML therapy.

摘要

由于据报道白细胞介素(IL-6)对急性髓性白血病(AML)原始细胞生长有多种作用,我们研究了原始细胞中IL-6受体(IL-6R)的表达水平是否与其对外源性IL-6增殖的敏感性相关。为了绝对定量IL-6R转录本数量,我们建立了一种带有内部RNA标准的定量IL-6R逆转录聚合酶链反应(RT-PCR)方法。在本研究中,研究了两种类型的AML原始细胞,即自主生长的细胞系细胞(n = 8)和未培养的天然原始细胞(n = 20),包括细胞系起源的那些细胞。天然原始细胞在一微克总细胞RNA中平均表达2.8×10⁷±1.9×10⁷个IL-6R转录本,而细胞系细胞的表达明显更丰富,值为8.3×10⁷±2.8×10⁷(P < 0.001)。通过在集落形成细胞培养试验中将细胞暴露于IL-6(1000 U/ml)来评估增殖反应,对于细胞系细胞,也在长期悬浮培养试验中进行评估。没有一个自主生长的细胞系对外源性IL-6有反应,而天然原始细胞表现出刺激、抑制或中性反应。因此,IL-6R表达水平不能预测细胞是否对外源性IL-6增殖,这表明在可能将这种细胞因子应用于AML治疗的临床使用之前,IL-6R定量不能用作筛选试验。

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