Original ContributionShikonin, a Chinese plant-derived naphthoquinone, induces apoptosis in hepatocellular carcinoma cells through reactive oxygen species: A potential new treatment for hepatocellular carcinoma
Graphical abstract
Highlights
► Shikonin, a naphthoquinone shows anti-cancer activity on hepatocellular carcinoma. ► Reactive Oxygen Species is involved in shikonin-induced apoptosis. ► Akt and RIP1/NF-κB signals are downstream of ROS generation. ► Results from animal experiments are consistent with in vitro studies. ► Shikonin may be a potential new treatment for hepatocellular carcinoma.
Section snippets
Cell lines and cell culture
The human hepatoma cell lines (HepG2, Huh7, and BEL7402) as well as the normal cell lines (L02 and HEK293T) were cultured in Dulbecco's modified Eagle's medium (DMEM) containing 10% fetal bovine serum, 100 units/ml penicillin, and 100 μg/ml streptomycin at 37 °C, 5% CO2.
Chemicals and antibodies
Shikonin was purchased from Enzo Life Sciences (Farmingdale, NY, USA). Z-VAD-fmk was from R&D Systems (Minneapolis, MN, USA), SP600125 was from Alexis Biochemicals (San Diego, CA, USA), LY294002 was from Cell Signaling Technology
Shikonin induces apoptosis in HCC cells but not in nontumorigenic cells
Although shikonin, the structure of which is shown in Fig. 1A, has been reported to induce cell death in various cancer cell lines and possesses anti-tumor activity in vivo, its action in HCC has not been studied in detail. To determine the effects of shikonin on HCC cells, dose-dependent response and time-course assessments of shikonin treatment were performed. Cell viability assays showed that shikonin induced cell death in Huh7, HepG2, and BEL7402 cells in a dose- and time-dependent manner.
Discussion
Taking into account the systemic side effects of cancer chemotherapy, the active ingredients of Chinese medical herbs are considered to be safe agents because of a long history of clinical applications in Chinese traditional medicine. Shikonin, a naphthoquinone derivative, is a key ingredient in traditional Chinese medicine that has been broadly used [47].
Many reports indicate that shikonin exerts multiple functions, including promoting wound healing with antibacterial and anti-inflammatory
Acknowledgments
This work was supported by the National Nature Science Foundation of China (30871261), the Chinese 111 Project (B06018), Fundamental Research Funds for the Central Universities (3081009), the Nature Science Foundation of Hubei Province (2008CDB080), the Program for Chenguang Young Scientists for Wuhan (200950431188), the National Basic Research Program of China (2010CB529800), and and the Project Sponsored by the Scientific Research Foundation for the Returned Overseas Chinese Scholars, State
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