Zuo Dongqing, Zhou Zifei, Wang Hongsheng, Zhang Tao, Zang Jie, Yin Fei, Sun Wei, Chen Jiepeng, Duan Lili, Xu Jing, Wang Zhuoying, Wang Chongren, Lin Binhui, Fu Zeze, Liao Yuxin, Li Suoyuan, Sun Mengxiong, Hua Yingqi, Zheng Longpo, Cai Zhengdong
Department of Orthopedics, Shanghai Tenth People's Hospital, School of Medicine, Tongji University, Shanghai, China.
Department of Orthopaedics, Shanghai First People's Hospital, Shanghai Jiao Tong University, Shanghai, China.
J Cell Mol Med. 2017 Feb;21(2):208-221. doi: 10.1111/jcmm.12957. Epub 2016 Sep 14.
Osteosarcoma (OS) is the most frequent primary malignant bone tumour. Alternol, a novel compound purified from microbial fermentation products exerts anti-tumour effects across several cancer types. The effect of alternol on human OS remains to be elucidated. We first evaluated the anti-tumour effect of alternol in several human OS cell lines in vitro and investigated its underlying mechanism. Alternol inhibited OS cell proliferation, migration and induced caspase-dependent apoptosis, G2/M cell cycle arrest in a dose and time-dependent manner. Moreover, alternol treatment inhibited signal transducer and activator of transcription-3 (STAT3) phosphorylation in 143B and MG63 human OS cells, as evaluated using a STAT3-dependent dual luciferase reporter system. Exposure to alternol resulted in excessive reactive oxygen species (ROS) generation and Jun amino-terminal kinases (JNK), extracellular signal-regulated kinases (ERK1/2) and p38 activation. Furthermore, alternol-induced cell death was significantly restored in the presence of the ROS scavenger, N-acetyl-l-cysteine (NAC) or a caspase inhibitor Z-VAD-FMK. NAC also prevented G2/M phase arrest and phosphorylation of mitogen-activated protein kinases (MAPK), but did not reverse STAT3 inactivation. Finally, alternol suppressed tumour growth in vivo in the nude mouse OS tibia orthotopic model. Immunohistochemistry revealed that alternol treatment resulted in down-regulation of phosph-STAT3 Tyr705 and up-regulation of cleaved caspase-3 and phosph-SAPK (Stress-activated protein kinases)/JNK expression. Taken together, our results reveal that alternol suppresses cell proliferation, migration and induces apoptosis, cell cycle arrest by modulating of ROS-dependent MAPK and STAT3 signalling pathways in human OS cells. Therefore, alternol is a promising candidate for developing anti-tumour drugs target OS.
骨肉瘤(OS)是最常见的原发性恶性骨肿瘤。Alternol是一种从微生物发酵产物中纯化得到的新型化合物,对多种癌症类型均具有抗肿瘤作用。然而,Alternol对人骨肉瘤的作用仍有待阐明。我们首先在体外评估了Alternol对几种人骨肉瘤细胞系的抗肿瘤作用,并研究了其潜在机制。Alternol以剂量和时间依赖性方式抑制骨肉瘤细胞增殖、迁移并诱导半胱天冬酶依赖性凋亡、G2/M期细胞周期阻滞。此外,使用STAT3依赖性双荧光素酶报告系统评估发现,Alternol处理可抑制143B和MG63人骨肉瘤细胞中信号转导和转录激活因子3(STAT3)的磷酸化。暴露于Alternol会导致活性氧(ROS)过度生成以及Jun氨基末端激酶(JNK)、细胞外信号调节激酶(ERK1/2)和p38激活。此外,在存在ROS清除剂N-乙酰-L-半胱氨酸(NAC)或半胱天冬酶抑制剂Z-VAD-FMK的情况下,Alternol诱导的细胞死亡得到显著恢复。NAC还可防止G2/M期阻滞和丝裂原活化蛋白激酶(MAPK)的磷酸化,但不能逆转STAT3失活。最后,在裸鼠骨肉瘤胫骨原位模型中,Alternol在体内抑制了肿瘤生长。免疫组织化学显示,Alternol处理导致磷酸化STAT3 Tyr705下调,以及裂解的半胱天冬酶-3和磷酸化应激激活蛋白激酶(SAPK)/JNK表达上调。综上所述,我们的结果表明,Alternol通过调节人骨肉瘤细胞中ROS依赖性MAPK和STAT3信号通路来抑制细胞增殖、迁移并诱导凋亡和细胞周期阻滞。因此,Alternol是开发针对骨肉瘤的抗肿瘤药物的有前途的候选物。