Cancer Prevention and Treatment Institute of Chengdu, Department of Oncology, Chengdu Fifth People's Hospital (The Second Clinical Medical College, Affiliated Fifth People's Hospital of Chengdu University of Traditional Chinese Medicine), Chengdu, 611137, China.
Department of Orthopedics, Haian traditional Chinese Medical Hospital, Haian Jiangsu, 226600, China.
Acta Biochim Pol. 2022 Aug 17;69(3):543-549. doi: 10.18388/abp.2020_5769.
Osteosarcoma, a leading malignant tumor of bones is diagnosed mostly in adolescents and young adults worldwide. The present study investigated alstonine as anti-osteosarcoma agent in vitro as well as in vivo and evaluated the underlying mechanism. Treatment with alstonine led to a significant (P<0.05) reduction in MG63 and U-2OS cell viability. Alstonine treatment of MG63 and U-2OS cells caused a significant reduction in colony formation compared to the control cells. Viability of osteoblasts was not affected by alstonine treatment in 1.25 to 20 µM concentration range. In alstonine treated MG63 and U-2OS cells apoptotic cells increased significantly (P<0.05) compared to the control cells. Moreover, in MG63 and U-2OS cells treatment with alstonine caused a prominent increase in expression of cleaved caspase-9, caspase-3, and PARP. Treatment of MG63 and U-2OS cells with alstonine caused a prominent increase in AMPKα (Thr172) phosphorylation and elevated the count of mtDNA copies compared to the untreated cells. Alstonine treatment of the cells caused a remarkable increase in expression of PGC-1α and TFAM proteins. Treatment of the mice with alstonine at 5 and 10 mg/kg doses for 30 days caused a significant (P<0.05) reduction in xenograft growth. Expression of PGC-1α and TFAM proteins in tumor tissues of the mice treated with alstonine was significantly (P<0.05) raised compared to the control group. Thus, alstonine inhibits osteosarcoma cell growth and activates apoptosis through AMPK dependent pathway. Therefore, alstonine may be considered for treatment of osteosarcoma as it effectively arrests tumor growth in mice.
骨肉瘤是一种主要发生在全球青少年和年轻人中的骨恶性肿瘤。本研究在体外和体内研究了 alstonine 作为抗骨肉瘤药物，并评估了其潜在机制。用 alstonine 治疗可显著（P<0.05）降低 MG63 和 U-2OS 细胞活力。与对照细胞相比，alstonine 处理 MG63 和 U-2OS 细胞导致集落形成显著减少。在 1.25 至 20 µM 浓度范围内，alstonine 处理不会影响成骨细胞的活力。在 alstonine 处理的 MG63 和 U-2OS 细胞中，与对照细胞相比，凋亡细胞显著增加（P<0.05）。此外，在 MG63 和 U-2OS 细胞中，用 alstonine 处理导致 cleaved caspase-9、caspase-3 和 PARP 的表达明显增加。与未处理的细胞相比，用 alstonine 处理 MG63 和 U-2OS 细胞会导致 AMPKα（Thr172）磷酸化明显增加，并增加 mtDNA 拷贝数。用 alstonine 处理细胞会导致 PGC-1α 和 TFAM 蛋白的表达显著增加。用 5 和 10 mg/kg 剂量的 alstonine 处理小鼠 30 天可显著（P<0.05）降低异种移植物生长。与对照组相比，用 alstonine 处理的小鼠肿瘤组织中 PGC-1α 和 TFAM 蛋白的表达显著（P<0.05）升高。因此，alstonine 通过 AMPK 依赖性途径抑制骨肉瘤细胞生长并激活细胞凋亡。因此，alstonine 可考虑用于治疗骨肉瘤，因为它可有效抑制小鼠肿瘤生长。
