Kohno N, Ohnuma T, Kawaida M, Ichikawa G
Department of Otolaryngology, Juntendo University School of Medicine, Tokyo, Japan.
Auris Nasus Larynx. 1993;20(1):53-62. doi: 10.1016/s0385-8146(12)80211-2.
We evaluated the effects of each component, natural human tumor necrosis factor-alpha (nHuTNF-alpha) and natural human interferon-alpha (nHu-IFN-alpha), and the combined nHuTNF-alpha/nHuIFN-alpha preparation with or without cisplatin (DDP) at tumor mass level. Multicellular tumor spheroids (MTS) of approximately 500 microns in diameter were produced from HEp-2 laryngeal squamous carcinoma cell line by liquid overlay culture technique. Cell kill effects of 72 hr exposure to nHuTNF-alpha, nHuIFN-alpha, or nHuTNF-alpha/nHuIFN-alpha against cells in MTS were 2-3-fold less than those in monolayer. In MTS, does response curves become progressively flat at high drug concentrations, indicative of poor drug penetration into the MTS core. Combination nHuTNF-alpha/nHuIFN-alpha produced synergistic cell kill for both MTS and monolayers. For monolayer cells exposure to nHuTNF-alpha/nHuIFN-alpha first (72 hr) followed by DDP (1 hr) after 1 hr rest period was synergistic with combination index (CI) of approximately 0.8 at LD50. Simultaneous (72 hr in nHuTNF-alpha/nHuIFN-alpha with DDP in last 1 hr) or reverse order of exposure were antagonistic. In contrast, for MTS, DDP followed by nHuTNF-alpha/nHuIFN-alpha was most synergistic with CI of approximately 0.2. Simultaneous exposure or nHuTNF-alpha/nHuIFN-alpha followed by DDP showed synergism with CI of approximately 0.5 and 0.8, respectively. The improved efficacy of DDP followed by nHuTNF-alpha/nHuIFN-alpha sequence for MTS cells appears to be due to increased nHuTNF-alpha/nHuIFN-alpha penetration into the MTS core aided by DDP.
我们在肿瘤块水平评估了天然人肿瘤坏死因子-α（nHuTNF-α）、天然人干扰素-α（nHu-IFN-α）以及联合使用nHuTNF-α/nHuIFN-α制剂（加或不加顺铂（DDP））的效果。通过液体覆盖培养技术，从人喉鳞状癌细胞系HEp-2制备出直径约500微米的多细胞肿瘤球体（MTS）。nHuTNF-α、nHuIFN-α或nHuTNF-α/nHuIFN-α对MTS中细胞72小时暴露的细胞杀伤效果比单层细胞低2至3倍。在MTS中，高药物浓度下剂量反应曲线逐渐变平，表明药物向MTS核心的渗透较差。联合使用nHuTNF-α/nHuIFN-α对MTS和单层细胞均产生协同细胞杀伤作用。对于单层细胞，先暴露于nHuTNF-α/nHuIFN-α（72小时），休息1小时后再暴露于DDP（1小时）具有协同作用，在半数致死剂量（LD50）时联合指数（CI）约为0.8。同时暴露（nHuTNF-α/nHuIFN-α中72小时，最后1小时加DDP）或相反顺序的暴露具有拮抗作用。相比之下，对于MTS，DDP后接nHuTNF-α/nHuIFN-α最具协同作用，CI约为0.2。同时暴露或nHuTNF-α/nHuIFN-α后接DDP分别表现出协同作用，CI约为0.5和0.8。DDP后接nHuTNF-α/nHuIFN-α序列对MTS细胞疗效的提高似乎是由于DDP有助于nHuTNF-α/nHuIFN-α向MTS核心的渗透增加。
