Lu Peace Wun-Ang, Lin Renn-Chia, Yang Jia-Sin, Lu Eric Wun-Hao, Hsieh Yi-Hsien, Tsai Meng-Ying, Lu Ko-Hsiu, Yang Shun-Fa
Morrison Academy Taichung, Taichung 406, Taiwan.
Department of Orthopedics, Chung Shan Medical University Hospital, Taichung 402, Taiwan.
Pharmaceuticals (Basel). 2021 May 24;14(6):497. doi: 10.3390/ph14060497.
Osteosarcoma is the most common primary bone malignancy in teenagers and continues to confer a generally poor prognosis due to its highly metastatic potential. Poor solubility in water and instability of curcumin limits its bioavailability for use in the adjuvant situation to improve the prognosis and the long-term survival of patients with osteosarcoma. To further obtain information regarding the apoptosis induced by a new curcumin analog, GO-Y078, in human osteosarcoma cells, flow cytometric analysis, annexin V-FITC/PI apoptosis staining assay, human apoptosis array, and Western blotting were employed. GO-Y078 dose-dependently decreased viabilities of human osteosarcoma U2OS, MG-63, 143B, and Saos-2 cells and induced sub-G1 fraction arrest and apoptosis in U2OS and 143B cells. In addition to the effector caspase 3 and poly adenosine diphosphate-ribose polymerase, GO-Y078 significantly activated both initiators of extrinsic caspase 8 and intrinsic caspase 9, whereas cellular inhibitors of apoptosis 1 (cIAP-1) and X-chromosome-linked IAP (XIAP) in U2OS and 143B cells were significantly repressed. Moreover, GO-Y078 increased phosphorylation of extracellular signal-regulated protein kinases (ERK)1/2, c-Jun N-terminal kinases (JNK)1/2, and p38 in U2OS and 143B cells. Using inhibitors of JNK (JNK-in-8) and p38 (SB203580), GO-Y078's increases in cleaved caspases 8, 9, and 3 could be expectedly suppressed, but they could not be affected by co-treatment with the ERK inhibitor (U0126). Altogether, GO-Y078 simultaneously induces both apoptotic pathways and cell arrest in U2OS and 143B cells through activating JNK and p38 signaling and repressing IAPs. These findings contribute to a better understanding of the mechanisms responsible for GO-Y078's apoptotic effects on human osteosarcoma cells.
骨肉瘤是青少年中最常见的原发性骨恶性肿瘤，由于其具有高度转移潜能，预后通常较差。姜黄素水溶性差且不稳定，限制了其在辅助治疗中用于改善骨肉瘤患者预后和长期生存的生物利用度。为了进一步了解新型姜黄素类似物GO-Y078诱导人骨肉瘤细胞凋亡的相关信息，采用了流式细胞术分析、膜联蛋白V-FITC/PI凋亡染色检测、人凋亡芯片和蛋白质免疫印迹法。GO-Y078剂量依赖性地降低人骨肉瘤U2OS、MG-63、143B和Saos-2细胞的活力，并诱导U2OS和143B细胞出现亚G1期阻滞和凋亡。除了效应半胱天冬酶3和聚腺苷二磷酸核糖聚合酶外，GO-Y078还显著激活了外源性半胱天冬酶8和内源性半胱天冬酶9这两种起始因子，而U2OS和143B细胞中的细胞凋亡抑制因子1（cIAP-1）和X染色体连锁凋亡抑制蛋白（XIAP）则受到显著抑制。此外，GO-Y078增加了U2OS和143B细胞中细胞外信号调节蛋白激酶（ERK）1/2、c-Jun氨基末端激酶（JNK）1/2和p38的磷酸化。使用JNK抑制剂（JNK-in-8）和p38抑制剂（SB203580），可以预期GO-Y078诱导的半胱天冬酶8、9和3的裂解增加会受到抑制，但与ERK抑制剂（U0126）联合处理则不会对其产生影响。总之，GO-Y078通过激活JNK和p38信号通路并抑制IAPs，在U2OS和143B细胞中同时诱导凋亡途径和细胞阻滞。这些发现有助于更好地理解GO-Y078对人骨肉瘤细胞凋亡作用的机制。
