Yun Hyung-Mun, Kim Soo Hyun, Kwon Yoon-Ju, Park Kyung-Ran
Department of Oral and Maxillofacial Pathology, School of Dentistry, Kyung Hee University, Seoul 02447, Republic of Korea.
National Development Institute for Korean Medicine, Gyeongsan 38540, Republic of Korea.
Antioxidants (Basel). 2024 Sep 25;13(10):1162. doi: 10.3390/antiox13101162.
Osteosarcoma is a primary malignant tumor found in the bones of children and adolescents. Unfortunately, many patients do not respond well to treatment and succumb to the illness. Therefore, it is necessary to discover novel bioactive compounds to overcome therapeutic limitations. Wang et Tang is a well-known herb used in oriental medicine. Studies have shown that metabolic diseases can be clinically treated using the roots of . Recent studies have demonstrated the anticarcinoma potential of root extracts; however, the exact mechanism remains unclear. The aim of this study was to examine the anti-osteosarcoma activity of a single compound extracted from the dried roots of . We purified Spicatoside A (SpiA) from the dried roots of . SpiA significantly inhibited the proliferation of human osteosarcoma MG63 cells in a dose- and time-dependent manner. SpiA also regulated the expression of various downstream proteins that mediate apoptosis (PARP, Bcl-2, and Bax), cell growth (cyclin D1, Cdk4, and Cdk6), angiogenesis (VEGF), and metastasis (MMP13). The Proteome Profiler Human Phospho-Kinase Array Kit showed that the AKT signaling protein was a target of SpiA in osteosarcoma cells. We also found that SpiA suppressed the constitutive activation of the PI3K-AKT-mTOR-p70S6K1 signaling pathway. We further validated the effects of SpiA on the AKT signaling pathway. SpiA induced autophagosome formation and suppressed necroptosis (a form of programmed cell death). SpiA increased the generation of reactive oxygen species (ROS) and led to the loss of mitochondrial membrane potential. N-acetylcysteine (NAC)-induced inhibition of ROS generation reduced SpiA-induced AKT inhibition, apoptotic cell death, and anti-metastatic effects by suppressing cell migration and invasion. Overall, these results highlight the anti-osteosarcoma effect of SpiA by inhibiting the AKT signaling pathway through ROS generation, suggesting that SpiA may be a promising compound for the treatment of human osteosarcoma.
骨肉瘤是一种发生于儿童和青少年骨骼的原发性恶性肿瘤。不幸的是，许多患者对治疗反应不佳并最终因病去世。因此，有必要发现新型生物活性化合物以克服治疗上的局限性。王不留行是一种在东方医学中常用的草药。研究表明，其根部可用于临床治疗代谢性疾病。最近的研究已经证明了根部提取物的抗癌潜力；然而，确切机制仍不清楚。本研究的目的是检测从王不留行干燥根中提取的单一化合物的抗骨肉瘤活性。我们从王不留行干燥根中纯化出了穗花杉双黄酮A（SpiA）。SpiA以剂量和时间依赖性方式显著抑制人骨肉瘤MG63细胞的增殖。SpiA还调节了多种介导细胞凋亡（PARP、Bcl-2和Bax）、细胞生长（细胞周期蛋白D1、细胞周期蛋白依赖性激酶4和细胞周期蛋白依赖性激酶6）、血管生成（血管内皮生长因子）和转移（基质金属蛋白酶13）的下游蛋白的表达。蛋白质组分析人磷酸化激酶阵列试剂盒显示，AKT信号蛋白是SpiA在骨肉瘤细胞中的作用靶点。我们还发现SpiA抑制了PI3K-AKT-mTOR-p70S6K1信号通路的组成性激活。我们进一步验证了SpiA对AKT信号通路的影响。SpiA诱导自噬体形成并抑制坏死性凋亡（一种程序性细胞死亡形式）。SpiA增加了活性氧（ROS）的生成并导致线粒体膜电位丧失。N-乙酰半胱氨酸（NAC）诱导的ROS生成抑制通过抑制细胞迁移和侵袭降低了SpiA诱导的AKT抑制、凋亡细胞死亡和抗转移作用。总体而言，这些结果突出了SpiA通过ROS生成抑制AKT信号通路的抗骨肉瘤作用，表明SpiA可能是一种治疗人类骨肉瘤的有前景的化合物。
