背景:尽管采用了包括手术在内的多模式方法,化疗和放疗,横纹肌肉瘤(RMS)的5年无事件生存率,儿童时期最常见的软组织肉瘤,对于转移性患者来说仍然非常差,主要是由于肿瘤细胞的选择和增殖驱动抵抗机制。使用新药或靶向治疗与常规治疗相结合的基于个性化医学的方案有可能增强治疗效果。同时最大限度地减少各种人类恶性肿瘤对健康组织的损害,开始了几项临床试验。在这项研究中,我们分析了,第一次,合成d的复合物SFX-01的抗肿瘤活性,在α-环糊精中稳定的l-萝卜硫素(EvgenPharmaplc,英国),用作单剂并与辐照结合使用,在四种肺泡和胚胎RMS的临床前模型中。的确,SFX-01在临床前研究中显示了其调节参与炎症和氧化应激的细胞途径的能力的前景,这对于癌症治疗中的控制至关重要。
方法:RH30,RH4(肺泡RMS),使用RD和JR1(胚胎RMS)细胞系以及RMS的小鼠异种移植模型来评估SFX-01处理诱导的生物学和分子效应。流式细胞术和通过q-PCR和蛋白质印迹分析的关键标志物的调节用于评估细胞增殖,凋亡,暴露于SFX-01的RMS细胞中自噬和细胞内活性氧(ROS)的产生。还通过特定的测定研究了迁移和侵入的能力。在体外和体内研究中研究了SFX-01与电离辐射(IR)之间可能的协同作用。学生t检验或双向方差分析用于检验两个或多个比较的统计学显著性,分别。
结果:SFX-01治疗表现出细胞抑制和细胞毒性作用,由G2细胞周期阻滞介导,细胞凋亡诱导和自噬抑制。此外,SFX-01作为单一疗法并与IR组合能够抑制3D肿瘤球的形成和增殖。最后,SFX-01,当作为单一药物口服时,在RMS异种移植小鼠模型中显示出减少肿瘤块生长的功效;当与放射治疗方案结合使用时,观察到它协同行动,导致比单独添加每个暴露所预期的更积极的结果。
结论:总之,我们的结果为SFX-01在RMS肿瘤临床前模型中的抗肿瘤特性提供了证据,既作为独立治疗,也与照射相结合。这些即将到来的发现对于SFX-01抗RMS分子机制的深入研究以及在RMS患者中建立临床试验至关重要,以便使用SFX-01/IR共同治疗作为有前途的治疗方法。特别是在侵袭性RMS疾病的临床管理中。
BACKGROUND: Despite a multimodal approach including surgery, chemo- and radiotherapy, the 5-year event-free survival rate for rhabdomyosarcoma (RMS), the most common soft tissue sarcoma in childhood, remains very poor for metastatic patients, mainly due to the selection and proliferation of tumour cells driving resistance mechanisms. Personalised medicine-based protocols using new drugs or targeted therapies in combination with conventional treatments have the potential to enhance the therapeutic effects, while minimizing damage to healthy tissues in a wide range of human malignancies, with several clinical trials being started. In this study, we analysed, for the first time, the antitumour activity of SFX-01, a complex of synthetic d, l-
sulforaphane stabilised in alpha-cyclodextrin (Evgen Pharma plc, UK), used as single agent and in combination with irradiation, in four preclinical models of alveolar and embryonal RMS. Indeed, SFX-01 has shown promise in preclinical studies for its ability to modulate cellular pathways involved in inflammation and oxidative stress that are essential to be controlled in cancer treatment.
METHODS: RH30, RH4 (alveolar RMS), RD and JR1 (embryonal RMS) cell lines as well as mouse xenograft models of RMS were used to evaluate the biological and molecular effects induced by SFX-01 treatment. Flow cytometry and the modulation of key markers analysed by q-PCR and Western blot were used to assess cell proliferation, apoptosis, autophagy and production of intracellular reactive oxygen species (ROS) in RMS cells exposed to SFX-01. The ability to migrate and invade was also investigated with specific assays. The possible synergistic effects between SFX-01 and ionising radiation (IR) was studied in both the in vitro and in vivo studies. Student\'s t-test or two-way ANOVA were used to test the statistical significance of two or more comparisons, respectively.
RESULTS: SFX-01 treatment exhibited cytostatic and cytotoxic effects, mediated by G2 cell cycle arrest, apoptosis induction and suppression of autophagy. Moreover, SFX-01 was able to inhibit the formation and the proliferation of 3D tumorspheres as monotherapy and in combination with IR. Finally, SFX-01, when orally administered as single agent, displayed a pattern of efficacy at reducing the growth of tumour masses in RMS xenograft mouse models; when combined with a radiotherapy regime, it was observed to act synergistically, resulting in a more positive outcome than would be expected by adding each exposure alone.
CONCLUSIONS: In summary, our results provide evidence for the antitumour properties of SFX-01 in preclinical models of RMS tumours, both as a standalone treatment and in combination with irradiation. These forthcoming findings are crucial for deeper investigations of SFX-01 molecular mechanisms against RMS and for setting up clinical trials in RMS patients in order to use the SFX-01/IR co-treatment as a promising therapeutic approach, particularly in the clinical management of aggressive RMS disease.