Cytotoxic activity of difluoromethylornithine compared with fenretinide in neuroblastoma cell lines.
Cytotoxic activity of difluoromethylornithine compared with fenretinide in neuroblastoma cell lines.
复制标题
在神经母细胞瘤细胞系中，二氟甲基鸟氨酸与芬维A胺的细胞毒性活性比较。
DOI：
10.1002/pbc.27447
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发表时间：
2018
影响因子：
3.2
通讯作者：
Reynolds,CPatrick
中科院分区：
文献类型：
--
作者：
Makena,MonishR;Cho,HwangEui;Nguyen,ThinhH;Koneru,Balakrishna;Verlekar,DatteshU;Hindle,Ashly;Kang,MinH;Reynolds,CPatrick
背景13-cis-维甲酸和免疫疗法（在强化细胞毒性治疗完成后给予）维持治疗可改善高危神经母细胞瘤患者的预后。在早期临床试验中，合成类维生素A芬维A胺（4-HPR）在复发/难治性神经母细胞瘤中实现了多次完全缓解，全身毒性低，已被考虑用于维持治疗临床试验。二氟甲基鸟氨酸（DFMO，一种不可逆的鸟氨酸脱羧酶抑制剂，单药临床应答数据极少）正用于神经母细胞瘤的维持治疗。我们评估了DFMO和芬维A胺在神经母细胞瘤细胞系中的细胞毒性活性。ProcedureWe使用DIMSCAN细胞毒性试验在骨髓水平缺氧（5%O2）中测试了16个神经母细胞瘤细胞系。多胺测定的HPLC-质谱和细胞凋亡转移酶dUTP缺口末端标记（TUNEL）使用flowcytometer.ResultsAt临床可达到的水平（100 μM），DFMO显着降低（P< 0.05）多胺腐胺和实现适度的细胞毒性（<1 log（90%的细胞毒性）。长时间暴露（7天）或在2%和20%O2中培养不会增强DFMO的细胞毒性。然而，芬维A胺（10 μM）即使在低于神经母细胞瘤患者临床可达到的浓度（20 μM）时，也在14个细胞系中诱导≥ 1 log细胞杀伤。芬维A胺的平均IC_（90）和IC_（99）分别为4.7 ± 1 μM和9.9 ± 1.8 μM。DFMO没有诱导凋亡的显著增加（P> 0.05）（TUNEL法）。芬维A胺诱导的细胞凋亡明显高于DFMO或对照组（P< 0.001）。
BackgroundMaintenance therapy with 13‐cis‐retinoic acid and immunotherapy (given after completion of intensive cytotoxic therapy) improves outcome for high‐risk neuroblastoma patients. The synthetic retinoid fenretinide (4‐HPR) achieved multiple complete responses in relapse/refractory neuroblastoma in early‐phase clinical trials, has low systemic toxicity, and has been considered for maintenance therapy clinical trials. Difluoromethylornithine (DFMO, an irreversible inhibitor of ornithine decarboxylase with minimal single‐agent clinical response data) is being used for maintenance therapy of neuroblastoma. We evaluated the cytotoxic activity of DFMO and fenretinide in neuroblastoma cell lines.ProcedureWe tested 16 neuroblastoma cell lines in bone marrow‐level hypoxia (5% O2) using the DIMSCAN cytotoxicity assay. Polyamines were measured by HPLC–mass spectrometry and apoptosis by transferase dUTP nick end labeling (TUNEL) using flow cytometry.ResultsAt clinically achievable levels (100 μM), DFMO significantly decreased (P< 0.05) polyamine putrescine and achieved modest cytotoxicity (<1 log (90% cytotoxicity). Prolonged exposures (7 days) or culture in 2% and 20% O2did not enhance DFMO cytotoxicity. However, fenretinide (10 μM) even at a concentration lower than clinically achievable in neuroblastoma patients (20 μM) induced ≥ 1 log cell kill in 14 cell lines. The average IC90and IC99of fenretinide was 4.7 ± 1 μM and 9.9 ± 1.8 μM, respectively. DFMO did not induce a significant increase (P> 0.05) in apoptosis (TUNEL assay). Apoptosis by fenretinide was significantly higher (P< 0.001) compared with DFMO or controls.ConclusionsDFMO as a single agent has minimal cytotoxic activity for neuroblastoma cell lines.