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.
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。