Experimental Study of Hepatocellular Carcinoma Treatment by Shikonin Through Regulating PKM2

Abstract

Tong Liu, 1–3,* Sainan Li, 2,* Liwei Wu, 2 Qiang Yu, 2, 4 Jingjing Li, 1 Jiao Feng, 2 Jie Zhang, 2, 4 Jiaojiao Chen, 2, 4 Yuting Zhou, 2, 4 Jie Ji, 2 Kan Chen, 2 Yuqing Mao, 5 Fan Wang, 6 Weiqi Dai, 7 Xiaoming Fan, 8 Jianye Wu, 1 Chuanyong Guo 1, 2 1Department of Gastroenterology, Putuo People’s Hospital, Tongji University School of Medicine, Shanghai 200060, People’s Republic of China; 2Department of Gastroenterology, Shanghai Tenth People’s Hospital, Tongji University School of Medicine, Shanghai 200072, People’s Republic of China; 3Department of Gastroenterology, Shandong Provincial Hospital of Shandong University, Ji’nan 250000, People’s Republic of China; 4Department of Gastroenterology, Shanghai Tenth People’s Hospital, School of Clinical Medicine of Nanjing Medical University, Shanghai 200072, People’s Republic of China; 5Department of Gerontology, Shanghai General Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200080, People’s Republic of China; 6Department of Oncology, Shanghai General Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200080, People’s Republic of China; 7Department of Gastroenterology, Shanghai Institute of Liver Diseases, Zhongshan Hospital of Fudan University, Shanghai 200032, People’s Republic of China; 8Department of Gastroenterology, Jinshan Hospital of Fudan University, Jinshan, Shanghai 201508, People’s Republic of China*These authors contributed equally to this workCorrespondence: Jianye WuDepartment of Gastroenterology, Putuo People’s Hospital, Tongji University School of Medicine, Shanghai 200060, People’s Republic of ChinaEmail [email protected] GuoDepartment of Gastroenterology, Shanghai Tenth People’s Hospital, Tongji University School of Medicine, Shanghai 200072, People’s Republic of ChinaEmail [email protected]: Shikonin is a natural product with many activities, including anti-cancer effects. Pyruvate kinase type M2 (PKM2) plays a crucial role in the growth of tumor cells. However, the effect of shikonin on PKM2 in hepatocellular carcinoma (HCC) is unclear.Methods: Cell viability, apoptosis level, glucose uptake, and lactate production were detected in HCC cells. Lentivirus-overexpressed and –shRNA of PKM2 were used to verify the key target of shikonin. A xenograft mouse model was used to detect the efficacy of shikonin and its combination with sorafenib in vivo.Results: Shikonin inhibited proliferation and glycolysis and induced apoptosis in HCC cells. Either PKM2-overexpressed or PKM2-shRNA alleviated or enhanced this effect. The results of CCK-8 showed that shikonin significantly inhibited cell viability of HCC cells. The levels of glucose uptake and lactate production were dramatically decreased by shikonin-treated. Results of flow cytometry and Western blot showed that the levels of apoptosis of HCC cells were significantly increased in a dose-dependent manner after shikonin treatment. In addition, shikonin enhanced the anti-cancer effect of sorafenib in vitro and in vivo. Our results showed that SK combined with sorafenib markedly inhibits tumor growth in HCC-transplanted nude mice compared to SK or sorafenib alone.Conclusion: By inhibiting PKM2, shikonin inhibited proliferation and glycolysis and induced cell apoptosis in HCC cells. The effect of shikonin on tumor cell proliferation, apoptosis and glycolsis will make it promising drug for HCC patients.Keywords: shikonin, PKM2, glycolysis, apoptosis, proliferation, hepatocellular carcinoma

Keywords

• shikonin
• pkm2
• glycolysis
• apoptosis
• proliferation
• hepatocellular carcinoma