Rutka J T, Giblin J R, Berens M E, Bar-Shiva E, Tokuda K, McCulloch J R, Rosenblum M L, Eessalu T E, Aggarwal B B, Bodell W J
Department of Neurological Surgery, School of Medicine, University of California, San Francisco 94143.
Int J Cancer. 1988 Apr 15;41(4):573-82. doi: 10.1002/ijc.2910410417.
To determine whether tumor necrosis factor is of potential value for the treatment of human malignant gliomas, we studied the effects of human recombinant tumor necrosis factor (rTNF-alpha) on the morphology, incorporation of tritiated thymidine, and proliferation of 5 established cell lines derived from human malignant gliomas and 3 normal human brain cell cultures. A radioreceptor analysis for rTNF-alpha was performed on all cell lines and cultures. Two of the 5 human glioma cell lines (SF-188 and U 343 MG-A) demonstrated a marked decrease (60% or less of untreated controls) in the uptake of tritiated thymidine when treated with rTNF-alpha at a concentration of 40 U/ml; rTNF-alpha at 100 U/ml had antiproliferative and cytotoxic effects on both cell lines. The growth and proliferation of cell lines SF-126 and U 251 MG were not affected by rTNF-alpha even at high concentrations (5,000 U/ml). The growth and proliferation of SF-539 were affected to an intermediate degree. A colony-forming efficiency assay corroborated the results of the proliferation studies: SF-126 was relatively resistant (surviving fraction of 0.9 at 500 U/ml) and SF-188 was relatively sensitive (surviving fraction of 0.08 at 500 U/ml) to the cytotoxic effects of rTNF-alpha. Time-sequence electron microscopy showed that rTNF-alpha at a concentration of 500 U/ml caused ultrastructural changes in SF-188, including increased intracytoplasmic vesiculation, swelling and degeneration of mitochondria, loss of cell:cell junctional complexes, and fragmentation of the plasma membrane. Studies with 125I-rTNF-alpha showed a variable degree of binding in all cell lines and cultures. SF-188, a highly sensitive cell line, demonstrated the strongest binding of 125I-rTNF-alpha (3,400 receptors/cell with high affinity; kd = 0.27 nM), while SF-126, a highly resistant cell line, had the weakest binding (809 receptors/cell; kd = 0.25 nM). We conclude that there is a spectrum of antiproliferative and cytotoxic activity among glioma-derived tumor cell lines exposed to rTNF-alpha. An increased number of rTNF-alpha receptors appears to be a necessary but insufficient condition to explain the antiproliferative effects observed in some glioma-derived cell lines.
为了确定肿瘤坏死因子对于治疗人类恶性胶质瘤是否具有潜在价值，我们研究了重组人肿瘤坏死因子（rTNF-α）对5种源自人类恶性胶质瘤的已建立细胞系以及3种正常人类脑细胞培养物的形态、氚标记胸腺嘧啶核苷掺入及增殖的影响。对所有细胞系和培养物进行了rTNF-α的放射受体分析。5种人类胶质瘤细胞系中的两种（SF-188和U 343 MG-A），当用浓度为40 U/ml的rTNF-α处理时，氚标记胸腺嘧啶核苷的摄取显著减少（为未处理对照的60%或更低）；100 U/ml的rTNF-α对这两种细胞系均有抗增殖和细胞毒性作用。即使在高浓度（5000 U/ml）下，SF-126和U 251 MG细胞系的生长和增殖也不受rTNF-α影响。SF-539的生长和增殖受到中等程度的影响。集落形成效率分析证实了增殖研究的结果：SF-126相对耐药（500 U/ml时存活分数为0.9），而SF-188对rTNF-α的细胞毒性作用相对敏感（500 U/ml时存活分数为0.08）。时间序列电子显微镜显示，浓度为500 U/ml的rTNF-α引起SF-188超微结构改变，包括胞质内小泡增多、线粒体肿胀和变性、细胞间连接复合物丧失以及质膜断裂。用125I-rTNF-α进行的研究表明，在所有细胞系和培养物中结合程度各不相同。高度敏感细胞系SF-188对125I-rTNF-α的结合最强（高亲和力时为3400个受体/细胞；kd = 0.27 nM），而高度耐药细胞系SF-126的结合最弱（809个受体/细胞；kd = 0.25 nM）。我们得出结论，在暴露于rTNF-α的胶质瘤衍生肿瘤细胞系中存在一系列抗增殖和细胞毒性活性。rTNF-α受体数量增加似乎是解释在某些胶质瘤衍生细胞系中观察到的抗增殖作用的必要但不充分条件。
