Kuppen P J, Jonges L E, van de Velde C J, Vahrmeijer A L, Tollenaar R A, Borel Rinkes I H, Eggermont A M
Department of Surgery, Leiden University Hospital, The Netherlands.
Br J Cancer. 1997;75(10):1497-500. doi: 10.1038/bjc.1997.255.
In this study we determined the level of tumour necrosis factor alpha (TNF-alpha) in liver and tumour tissue samples obtained from patients with colorectal metastases confined to the liver, who were treated with isolated liver perfusion with TNF-alpha and melphalan. We adapted a standard enzyme-linked immunosorbent assay kit for the quantification of TNF-alpha in serum to measure the amount of this cytokine in solid tissue. For this purpose, we developed a buffer that lysed the tissues without affecting the TNF-alpha present. The minimum detection level was about 2 pg of TNF-alpha per mg tissue. Using this technique, we found a significant increase in the TNF-alpha level after perfusion in the liver tissue of all evaluable patients, which may explain the transient liver toxicity we observed in all patients. In tumour tissue, a significant TNF-alpha increase was observed in one out of five patients. The level of TNF-alpha in all liver tissue samples and some of the tumours after treatment by isolated liver perfusion was much higher than the peak serum concentrations obtained after systemic administration of the maximum tolerated dose of TNF-alpha. Furthermore, we demonstrated that the level of TNF-alpha in the liver tissue samples was about seven to eight times higher than in tumour tissue. We concluded that regional liver treatment resulted in a relatively high local level of TNF-alpha, but also that this cytokine did not preferentially accumulate in tumour tissue.
在本研究中，我们测定了从局限于肝脏的结直肠癌转移患者获取的肝脏和肿瘤组织样本中肿瘤坏死因子α（TNF-α）的水平，这些患者接受了TNF-α和美法仑的离体肝脏灌注治疗。我们采用一种用于定量血清中TNF-α的标准酶联免疫吸附测定试剂盒来测量实体组织中这种细胞因子的含量。为此，我们开发了一种能裂解组织而不影响其中TNF-α的缓冲液。最低检测水平约为每毫克组织2皮克TNF-α。使用该技术，我们发现所有可评估患者肝脏组织灌注后TNF-α水平显著升高，这可能解释了我们在所有患者中观察到的短暂性肝脏毒性。在肿瘤组织中，五分之一的患者观察到TNF-α显著升高。离体肝脏灌注治疗后，所有肝脏组织样本以及部分肿瘤中的TNF-α水平远高于全身给予最大耐受剂量TNF-α后获得的血清峰值浓度。此外，我们证明肝脏组织样本中的TNF-α水平比肿瘤组织中的高约7至8倍。我们得出结论，区域性肝脏治疗导致局部TNF-α水平相对较高，但这种细胞因子并非优先在肿瘤组织中蓄积。
