Dunn T A, Schmoll H J, Rie C, Hartmann K, Casper J
Department of Haematology and Oncology, University of Halle-Wittenberg, Halle/Saale, Germany.
J Cancer Res Clin Oncol. 1998;124(8):435-43. doi: 10.1007/s004320050196.
Peptide growth factors involved in the regulation of haematopoiesis (HPGF), for example granulocyte-colony-stimulating factor (G-CSF) and granulocyte/macrophage-colony-stimulating factor (GM-CSF), are of clinical importance in the treatment of testicular germ cell tumour (GCT) patients with modern chemotherapy regimens since they ameliorate chemotherapy-induced neutropenia. Aberrant expression of and/or response to HPGF has been reported in several solid tumour types although no data are available on GCT with the exception of those on stem cell factor (SCF). The aims of this pre-clinical study were twofold: (1) to screen a panel of human non-seminomatous (NS)GCT for the production of HPGF and (2) to test the effects of G-CSF or SCF on the growth of NSGCT cell lines in vitro, and on the growth kinetics of two human NSGCT xenograft models. HPGF concentrations in cell culture supernatant from 11 NSGCT cell lines growing under routine culture conditions were measured by enzyme-linked immunosorbent assay. The growth kinetics of cell lines was quantified in vitro using the sulphorhodamine B assay. The growth kinetics of nude mouse NSGCT xenografts was followed by measuring tumour volumes every 2-3 days over days 1-30, following daily subcutaneous injection of nude mice (days 1-14). The cell lines produced G-CSF (1/11 cell lines), GM-CSF (2/11), SCF (2/11), M-CSF (6/11). and interleukin-6 (9/11). Growth stimulation of cell line H12.1 by SCF was observed in vitro, but no statistically significant differences in NSGCT xenograft tumour volume (VT) or relative VT (r VT) in treated groups were observed on days 14 or 29 compared to the control. The change in rVT of H12.1 xenografts treated with G-CSF alone compared to control (rVT/rVT,c) was 0.96 on day 29. The values for rVT/rVT,c for H12.1 xenografts treated with G-CSF in combination with low- or high-dose SCF were, respectively, 1.67 or 1.7 compared to 1.19 for SCF-treated mice. The results are in agreement with clinical data to date where no observations have been reported of stimulation or inhibition of tumours in patients receiving treatment with G-CSF. Before any clinical trials are initiated in GCT patients treated with G-CSF in combination with SCF, further pre-clinical experiments with this tumour type are recommended to investigate this phenomenon further in a greater number of NSGCT cell lines in vitro and in vivo and with a wider range of SCF/G-CSF schedules. The potential relevance of secretion of HPGF in NSGCT cell lines in vitro to the pathobiology of GCT in patients is also a subject of interest for future research.
参与造血调节的肽生长因子（HPGF），例如粒细胞集落刺激因子（G-CSF）和粒细胞/巨噬细胞集落刺激因子（GM-CSF），在采用现代化疗方案治疗睾丸生殖细胞肿瘤（GCT）患者时具有临床重要性，因为它们可改善化疗引起的中性粒细胞减少。尽管除了关于干细胞因子（SCF）的数据外，尚无GCT的相关数据，但已有报道称在几种实体瘤类型中存在HPGF的异常表达和/或反应。这项临床前研究的目的有两个：（1）筛选一组人类非精原细胞瘤（NS）GCT以检测HPGF的产生；（2）测试G-CSF或SCF对NSGCT细胞系体外生长以及两个人类NSGCT异种移植模型生长动力学的影响。通过酶联免疫吸附测定法测量在常规培养条件下生长的11个NSGCT细胞系的细胞培养上清液中的HPGF浓度。使用磺基罗丹明B测定法在体外对细胞系的生长动力学进行定量。在裸鼠每日皮下注射（第1 - 14天）后的第1 - 30天，每2 - 3天测量一次肿瘤体积，以跟踪裸鼠NSGCT异种移植瘤的生长动力学。这些细胞系产生G-CSF（1/11个细胞系）、GM-CSF（2/11）、SCF（2/11）、M-CSF（6/11）和白细胞介素-6（9/11）。在体外观察到SCF对细胞系H12.1有生长刺激作用，但与对照组相比，在第14天或第29天，治疗组的NSGCT异种移植瘤体积（VT）或相对VT（rVT）没有统计学上的显著差异。与对照组相比，仅用G-CSF治疗的H12.1异种移植瘤在第29天的rVT变化（rVT/rVT,c）为0.96。与SCF治疗的小鼠的1.19相比，用G-CSF联合低剂量或高剂量SCF治疗的H12.1异种移植瘤的rVT/rVT,c值分别为1.67或1.7。这些结果与迄今为止的临床数据一致，在接受G-CSF治疗的患者中尚未有肿瘤受到刺激或抑制的观察报告。在对接受G-CSF联合SCF治疗的GCT患者启动任何临床试验之前，建议对这种肿瘤类型进行进一步的临床前实验，以在更多的NSGCT细胞系中进行体外和体内研究，并采用更广泛的SCF/G-CSF给药方案进一步研究这一现象。NSGCT细胞系体外分泌HPGF与患者GCT病理生物学的潜在相关性也是未来研究感兴趣的课题。
