Shikonin-induced necroptosis is enhanced by the inhibition of autophagy in non-small cell lung cancer cells

• Published in: Journal of translational medicine Vol. 15; no. 1; p. 123
• Main Authors: Kim, Hyo-Jin, Hwang, Ki-Eun, Park, Do-Sim, Oh, Seon-Hee, Jun, Hong Young, Yoon, Kwon-Ha, Jeong, Eun-Taik, Kim, Hak-Ryul, Kim, Young-Suk
• Format: Journal Article
• Language: English
• Published: England BioMed Central Ltd 31-05-2017; BioMed Central; BMC
• Subjects: A549 Cells; Animal models; Animals; Annexin V; Antitumor activity; Apoptosis; Assaying; Autophagy; Autophagy (Cytology); Bafilomycin A; Carcinoma, Non-Small-Cell Lung - drug therapy; Carcinoma, Non-Small-Cell Lung - metabolism; Care and treatment; Caspase; Caspase 8 - metabolism; Caspase-8; Cell death; Cell Line, Tumor; Chinese herbal medicine; Chinese medicine; Cytometry; Cytotoxicity; Diagnosis; Gene Silencing; Humans; Imidazoles - pharmacology; Immunoglobulins; Indoles - pharmacology; Inhibition; Inhibitors; Kinases; Leukemia; Lithospermum; Lung cancer; Lung Neoplasms - drug therapy; Lung Neoplasms - metabolism; Macrolides - pharmacology; Mice; Mice, Inbred BALB C; Mice, Nude; Naphthoquinones - pharmacology; Necroptosis; Necrosis; Neoplasm Transplantation; Non-small cell lung cancer; Non-small cell lung carcinoma; Penicillin; Phagocytosis; RIP1; RNA, Small Interfering - metabolism; Shikonin; siRNA; Staining; Tissues; Western blotting; X-Ray Microtomography; Xenografts; United States > US; Necroptosis; Shikonin; Autophagy; RIP1; Non-small cell lung cancer
• ISSN: 1479-5876; 1479-5876
• DOI: 10.1186/s12967-017-1223-7

Shikonin, a natural naphthoquinone pigment purified from Lithospermum erythrorhizon, induces necroptosis in various cancer types, but the mechanisms underlying the anticancer activity of shikonin in lung cancer are not fully understood. This study was designed to clarify whether shikonin causes necroptosis in non-small cell lung cancer (NSCLC) cells and to investigate the mechanism of action.
Multiplex and caspase 8 assays were used to analyze effect of shikonin on A549 cells. Cytometry with annexin V/PI staining and MTT assays were used to analyze the mode of cell death. Western blotting was used to determine the effect of shikonin-induced necroptosis and autophagy. Xenograft and orthotopic models with A549 cells were used to evaluate the anti-tumor effect of shikonin in vivo.
Most of the cell death induced by shikonin could be rescued by the specific necroptosis inhibitor necrostatin-1, but not by the general caspase inhibitor Z-VAD-FMK. Tumor growth was significantly lower in animals treated with shikonin than in the control group. Shikonin also increased RIP1 protein expression in tumor tissues. Autophagy inhibitors, including methyladenine (3-MA), ATG5 siRNA, and bafilomycin A, enhanced shikonin-induced necroptosis, whereas RIP1 siRNA had no effect on the apoptotic potential of shikonin.
Our data indicated that shikonin treatment induced necroptosis and autophagy in NSCLC cells. In addition, the inhibition of shikonin-induced autophagy enhanced necroptosis, suggesting that shikonin could be a novel therapeutic strategy against NSCLC.