Berdel W E, Danhauser-Riedl S, Steinhauser G, Rastetter J
Department of Medicine I, Technische Universität München.
Onkologie. 1990 Dec;13(6):437-43. doi: 10.1159/000216816.
We studied the influence of recombinant human (rh) interleukin-3 (IL-3) and rh granulocyte-macrophage colony-stimulating factor (GM-CSF) on the clonal growth of a human colorectal adenocarcinoma cell line in a methylcellulose assay for colony growth of solid tumor cell lines (HTCAMC) and a capillary modification of a human tumor cloning assay in agar (HTCAcap). Both growth factors stimulated the clonal growth of this cell line in a dose-dependent fashion. Neutralizing the monoclonal antibody abolished the effect of rhGM-CSF. There was an inverse correlation between the spontaneous plating efficacy (PE) of the cells and their susceptibility to the stimulation by the growth factors. From day 4 to 7 we found conditions in which clusters and colonies occurred preferentially in the growth factor-stimulated cultures. Single colonies taken from these cultures grew rapidly into macroscopically visible tumors in liquid cultures and had a high secondary PE (PE2) in the HTCAcap, both presenting an argument against a differentiating effect of the growth factors on this tumor cell line. Furthermore, we were able to define conditions in which rhGM-CSF significantly increased the cytotoxicity of 5-fluorouracil (5-FU). However, this effect was dependent on spontaneous PE of the cells, degree of stimulation by the factor, degree of cytotoxicity of 5-FU in the controls, as well as the therapeutic regimen. Since there were only narrow margins for a beneficial effect of rhGM-CSF in this setting when absolute numbers of surviving colonies were counted, it remains doubtful whether this approach will be exploitable in the clinical situation.
我们在用于实体瘤细胞系集落生长的甲基纤维素测定法（HTCAMC）以及琼脂中人肿瘤克隆测定法的毛细管改良法（HTCAcap）中，研究了重组人（rh）白细胞介素-3（IL-3）和rh粒细胞-巨噬细胞集落刺激因子（GM-CSF）对人结肠直肠腺癌细胞系克隆生长的影响。两种生长因子均以剂量依赖性方式刺激该细胞系的克隆生长。中和性单克隆抗体消除了rhGM-CSF的作用。细胞的自发接种效率（PE）与其对生长因子刺激的敏感性之间呈负相关。在第4天至第7天，我们发现了一些条件，在此条件下，生长因子刺激的培养物中优先出现细胞团和集落。从这些培养物中取出的单个集落在液体培养物中迅速生长为肉眼可见的肿瘤，并且在HTCAcap中具有较高的二次接种效率（PE2），这两者都表明生长因子对该肿瘤细胞系没有分化作用。此外，我们能够确定rhGM-CSF可显著增加5-氟尿嘧啶（5-FU）细胞毒性的条件。然而，这种作用取决于细胞的自发PE、因子的刺激程度、对照中5-FU的细胞毒性程度以及治疗方案。由于在这种情况下，当计算存活集落的绝对数量时，rhGM-CSF产生有益作用的余地很小，因此这种方法在临床情况下是否可利用仍值得怀疑。
