The anti-tumor effect of shikonin on osteosarcoma by inducing RIP1 and RIP3 dependent necroptosis

BMC Cancer

Zeze Fu¹  Biyong Deng¹  Yuxin Liao¹  Liancheng Shan²  Fei Yin¹  Zhuoying Wang¹  Hui Zeng¹  Dongqing Zuo¹  Yingqi Hua¹  Zhengdong Cai¹ 

[1] Department of Orthopedics, Shanghai Tenth People’s Hospital, Tongji University, School of Medicine, Shanghai 200072, China

[2] Postdoctoral Research Station of School of Life Science and Technology of Tongji University, Shanghai 200092, China

关键词: RIP3;    RIP1;    Metastasis;    Shikonin;    Necroptosis;    Osteosarcoma;   

Others  :  859218
DOI  :  10.1186/1471-2407-13-580

 received in 2013-09-02, accepted in 2013-12-04,  发布年份 2013

【 摘 要 】

Background

Osteosarcoma is the most frequent primary malignant bone tumor, notorious for its lung metastasis. Shikonin, an effective constituent extracted from Chinese medicinal herb, was demonstrated to induce necroptosis in some cancers.

Methods

MTT assay was performed to detect cell survival rate in vitro. Flow cytometry was used to analyze cell cycle and cell death. Western blot was performed to determine the expression levels of RIP1, RIP3, caspase-3, caspase-6 and PARP. The tibial primary and lung metastatic osteosarcoma models were used to evaluate the anti-tumor effect of shikonin in vivo.

Results

The cell survival rate was decreased in a dose and time dependent manner when treated with shikonin. No major change in cell cycle was observed after shikonin treatment. The cell death induced by shikonin could be mostly rescued by specific necroptosis inhibitor necrostatin-1, but not by general caspase inhibitor Z-VAD-FMK. The number of necrotic cells caused by shikonin was decreased after being pretreated with Nec-1 detected by flow cytometry in K7 cells. After 8-hour treatment of shikonin, the expression levels of RIP1 and RIP3 were increased while caspase-3, caspase-6 and PARP were not activated in K7 and U2OS cells determined by Western blot. Size of primary tumor and lung metastasis in shikonin treated group were significantly reduced. The protein levels of RIP1 and RIP3 in primary tumor tissues were increased by shikonin. The overall survival of lung metastatic models was longer compared with control group (p < 0.001).

Conclusions

Shikonin had prompt but profound anti-tumor effect on both primary and metastatic osteosarcoma, probably by inducing RIP1 and RIP3 dependent necroptosis. Shikonin would be a potential anti-tumor agent on the treatment of primary and metastatic osteosarcoma.

【 授权许可】

   
2013 Fu et al.; licensee BioMed Central Ltd.

【 图 表 】

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