Komdeur R, Meijer C, Van Zweeden M, De Jong S, Wesseling J, Hoekstra H J, van der Graaf W T A
Department of Surgical Oncology, University Hospital of Groningen, 9700 RB Groningen, The Netherlands.
Int J Oncol. 2004 Sep;25(3):677-84. doi: 10.3892/ijo.25.3.677.
Doxorubicin (DOX) and ifosfamide (IFO) are the most active single agents in soft tissue sarcomas (STS). Tumour necrosis factor-alpha (TNF-alpha) is used for STS in the setting of isolated limb perfusions. Like TNF-alpha, TNF-related apoptosis-inducing ligand (TRAIL) induces apoptosis. In contrast to TNF-alpha preliminary studies suggest that TRAIL lacks systemic side effects. The effects of TRAIL alone and in combination with DOX or 4-hydroxy-IFO were evaluated in the TNF-alpha sensitive rhabdomyosarcoma cell line KYM-1, its 5-fold TNF-alpha sensitive subline KD4 and its >150-fold TNF-alpha resistant subline 37B8R. Membrane expression of TRAIL-receptors DR4 (death receptor 4), DR5 (pro-apoptotic), DcR1 (decoy receptor 1), DcR2 (anti-apoptotic) was assessed by flow cytometry. Cytotoxicity was determined by microculture tetrazolium assays. Apoptosis assays were performed with acridine orange. DOX (doxorubicin) and 4-OH-IFO decreased survival in all cell lines; a 2-fold resistance was observed for both drugs in 37B8R. All cell lines expressed DR4 and DR5, but hardly any DcR1 or DcR2. TRAIL was cytotoxic in KYM-1, even more in KD4 and induced massive apoptosis; 37B8R was >500-fold resistant to TRAIL and little apoptosis could be observed. TRAIL plus DOX showed synergistic cytotoxicity in KYM-1 and 37B8R. TRAIL plus 4-OH-IFO showed addition in all three cell lines. DOX plus TRAIL-induced more cytotoxicity and apoptosis in all cell lines compared to TRAIL alone. In 37B8R, DOX overcame resistance to TRAIL. In KYM-1, KD4 and 37B8R, sensitivity and resistance to TNF-alpha and TRAIL parallels. TRAIL-resistance was independent from expression of TRAIL-receptors. DOX with TRAIL could overcome TRAIL-resistance in 37B8R cells.
阿霉素（DOX）和异环磷酰胺（IFO）是软组织肉瘤（STS）中最有效的单一药物。肿瘤坏死因子-α（TNF-α）用于肢体隔离灌注治疗STS。与TNF-α一样，肿瘤坏死因子相关凋亡诱导配体（TRAIL）也可诱导凋亡。与TNF-α不同的是，初步研究表明TRAIL缺乏全身副作用。在对TNF-α敏感的横纹肌肉瘤细胞系KYM-1、其对TNF-α敏感5倍的亚系KD4及其对TNF-α耐药>150倍的亚系37B8R中，评估了TRAIL单独使用以及与DOX或4-羟基异环磷酰胺联合使用的效果。通过流式细胞术评估TRAIL受体DR4（死亡受体4）、DR5（促凋亡）、DcR1（诱饵受体1）、DcR2（抗凋亡）的膜表达。通过微量培养四氮唑试验测定细胞毒性。用吖啶橙进行凋亡试验。DOX（阿霉素）和4-OH-IFO降低了所有细胞系的存活率；在37B8R中观察到这两种药物均有2倍的耐药性。所有细胞系均表达DR4和DR5，但几乎不表达DcR1或DcR2。TRAIL对KYM-1具有细胞毒性，对KD4的毒性更强，并诱导大量凋亡；37B8R对TRAIL的耐药性>500倍，几乎观察不到凋亡。TRAIL加DOX在KYM-1和37B8R中显示出协同细胞毒性。TRAIL加4-OH-IFO在所有三个细胞系中均显示相加作用。与单独使用TRAIL相比，DOX加TRAIL在所有细胞系中诱导更多的细胞毒性和凋亡。在37B8R中，DOX克服了对TRAIL的耐药性。在KYM-1、KD4和37B8R中，对TNF-α和TRAIL的敏感性和耐药性相似。TRAIL耐药性与TRAIL受体的表达无关。DOX与TRAIL可克服37B8R细胞中的TRAIL耐药性。
