Analysis of Growth Factor Dependent Proliferation of Malignant Gliomas ; A Study for Novel Therapeutic Mode Utilizing Biological Response Modifications.

在本研究中，我们研究了多种细胞因子和生物活性分子；恶性胶质瘤细胞产生这些生长因子，反之，外源因子对肿瘤细胞的影响。胶质母细胞瘤细胞产生IL-1α、IL-1β、IL-6、IL-8、IL-10、G-CSF、SCF、PGE2、成纤维细胞生长因子和PDGF。IL-1和肿瘤坏死因子对胶质母细胞瘤细胞代谢的影响最为显著。用四甲基偶氮唑盐比色法显示，即使在高剂量(256U·ml-1)，肿瘤坏死因子对胶质母细胞瘤细胞也没有明显的杀伤活性，但在较低剂量时可显著抑制集落形成和DNA合成。流式细胞仪检测显示，低剂量(10U/ml)的肿瘤坏死因子可刺激PGE2、锰超氧化物歧化酶、IL-6和IL-8的产生。肿瘤坏死因子使某些人胶质瘤细胞滞留在G0/G1期，导致随后的S期DNA合成减少，抑制了细胞增殖。在早期的I期临床试验中，肿瘤坏死因子对6例胶质母细胞瘤患者进行了脑内注射。在这项试验中，作者研究了局部注射肿瘤坏死因子后脑脊液和局部液中的体内免疫反应。检测到这些体液中的肿瘤坏死因子，半衰期为几小时。注射肿瘤坏死因子后，出现大量的白细胞迁移。中性粒细胞在8~12h达到高峰，然后是CD4+CD8-T细胞和CDIIb+CD13+CD14+单核细胞。脑脊液中IL-8活性与中性粒细胞迁移高峰同时相对应，脑脊液和/或脑脊液中IL-6、IL-1b和PGB2水平在注射肿瘤坏死因子后8~12h达到高峰。IL-2和干扰素均未检测到。肿瘤坏死因子可能通过其直接的细胞抑制作用和间接的免疫调节作用发挥抗肿瘤作用。

In this study, various cytokines and bioactive molecules were studied ; productions of these growth factors by malignant gllioma cells and inversely the effect of exogenous factors on the tumor cells. In particular, glioma patients were investigated.Human glioblastoma cells produced IL-1alpha, IL-1beta, IL-6, IL-8, IL-10, G-CSF, SCF, PGE2, FGF, and PDGF.Glioblastoma cells expressed IL-1 receptors and p55TNF receptor. IL-1 and TNF had most remarkable effect on metabolism of the glioblastoma cells. TNF stimulation on glioblastoma cells, even at a high dose (256U-ml), exhibited no remarkable cytocidal activity in MTT assay, but at lower doses significantly inhibited colony forming and DNA synthesis. TNF at a low dose (10U/ml) stimulated production of PGE2, Mn-superoxide dismutase, IL-6 and IL-8 by flow cytometry. TNF arrested certain human glioma cells in the G0/G1 phase resulting in reduction of DNA synthesis in the subsequent S phase, suppressing the proliferation assay.In an early Phase I clinical trial, TNF was administered intracranially to six parients bearing glioblastoma. In this trial, the author studied in vivo immunological responses in the cerebrospinal fluid and regional fluid after the regional TNF injections. TNF in these body fluid were detected with a half life of several hours. There occurred a substantial number of leukocyte migration after the TNF administration. Neutrophils appeared first peaking at 8 to 12 hours, and then CD4+CD8-T cells and CDIIb+CD13+CD14+ monocytes followed. IL-8 activity in the cerebrospinal fluid simultaneously corresponded to the peak of the neutrophil migration Increases in IL-6, IL-1b and PGB2 levels in the cerebrospinal fluid, regional fluid or both occurred peaking at 8 to 12 hours after TNF injection. Neither IL-2 nor interfarons were detected. TNF may act as an antineoplastic agent by its direct cytostatic effects and indirectly through immune modulatry effects.