Zorba Başak Işıl, Boyacıoğlu Özge, Çağlayan Tuğba, Reçber Tuba, Nemutlu Emirhan, Eroğlu İpek, Korkusuz Petek
Graduate School of Science and Engineering, Department of Bioengineering, Hacettepe University, Ankara, Turkey.
Faculty of Medicine, Department of Medical Biochemistry, Atılım University, Ankara, Turkey.
J Liposome Res. 2024 Jun;34(2):274-287. doi: 10.1080/08982104.2023.2262025. Epub 2023 Oct 5.
Curable approaches for primary osteosarcoma are inadequate and urge investigation of novel therapeutic formulations. Cannabinoid ligands exert antiproliferative and apoptotic effect on osteosarcoma cells cannabinoid 2 (CB2) or transient receptor potential vanilloid type (TRPV1) receptors. In this study, we confirmed CB2 receptor expression in MG63 and Saos-2 osteosarcoma cells by qRT-PCR and flow cytometry (FCM), then reported the reduction effect of synthetic specific CB2 receptor agonist CB65 on the proliferation of osteosarcoma cells by WST-1 (water-soluble tetrazolium-1) and RTCA (real-time impedance-based proliferation). CB65 revealed an IC50 (inhibitory concentration) for MG63 and Saos-2 cells as 1.11 × 10 and 4.95 × 10 M, respectively. The specific antiproliferative effect of CB65 on osteosarcoma cells was inhibited by CB2 antagonist AM630. CB65 induced late apoptosis of MG63 and Saos-2 cells at 24 and 48 h, respectively by FCM when applied submaximal concentration. A novel CB65 liposomal system was generated by a thin film hydration method with optimal particle size (141.7 ± 0.6 nm), polydispersity index (0.451 ± 0.026), and zeta potential (-10.9 ± 0.3 mV) values. The encapsulation efficiency (EE%) of the CB65-loaded liposomal formulation was 51.12%. The CB65 and CB65-loaded liposomal formulation releasing IC50 of CB65 reduced proliferation by RTCA and invasion by scratch assay and induced late apoptosis of MG63 and Saos-2 cells, by FCM. Our results demonstrate the CB2 receptor-mediated antiproliferative and apoptotic effect of a new liposomal CB65 delivery system on osteosarcoma cells that can be used as a targeted and intelligent tool for bone tumors to ameliorate pediatric bone cancers following validation.
原发性骨肉瘤的可治愈方法并不完善，因此迫切需要研究新的治疗方案。大麻素配体对骨肉瘤细胞、大麻素2（CB2）或瞬时受体电位香草酸亚型1（TRPV1）受体具有抗增殖和凋亡作用。在本研究中，我们通过qRT-PCR和流式细胞术（FCM）证实了MG63和Saos-2骨肉瘤细胞中CB2受体的表达，然后通过WST-1（水溶性四唑盐-1）和RTCA（基于实时阻抗的增殖）报告了合成特异性CB2受体激动剂CB65对骨肉瘤细胞增殖的抑制作用。CB65对MG63和Saos-2细胞的IC50（抑制浓度）分别为1.11×10和4.95×10 M。CB2拮抗剂AM630抑制了CB65对骨肉瘤细胞的特异性抗增殖作用。当以亚最大浓度应用时，CB65分别在24小时和48小时通过FCM诱导MG63和Saos-2细胞晚期凋亡。通过薄膜水化法制备了一种新型CB65脂质体系统，其具有最佳粒径（141.7±0.6 nm）、多分散指数（0.451±0.026）和zeta电位（-10.9±0.3 mV）值。负载CB65的脂质体制剂的包封率（EE%）为51.12%。CB65和负载CB65的脂质体制剂释放的CB65的IC使通过RTCA的增殖减少，通过划痕试验的侵袭减少，并通过FCM诱导MG63和Saos-2细胞晚期凋亡。我们的结果证明了新的脂质体CB65递送系统通过CB2受体介导的对骨肉瘤细胞的抗增殖和凋亡作用，该系统经验证后可作为骨肿瘤的靶向智能工具来改善儿童骨癌。
