Haibo Wang1, Lide Tao2, Tengyang Ni3, Hao Gu4, Feng Jin5, Xiaojun Dai6, Jun Feng7, Yanbing Ding8, Weiming Xiao9, Shiyu Guo10, Tadashi Hisamitsu11, Yayun Qian12, Yanqing Liu13. 1. The Affiliated Hospital of Yangzhou University, Yangzhou University, Yangzhou 225000, China; The State Administration of Traditional Chinese Medicine Key Laboratory of Toxic Pathogens-Based Therapeutic Approaches of Gastric Cancer, Yangzhou 225000, China; Clinical Medicine College of Yangzhou University, Yangzhou 225000, China. Electronic address: whbosy@163.com. 2. The Affiliated Hospital of Yangzhou University, Yangzhou University, Yangzhou 225000, China; The State Administration of Traditional Chinese Medicine Key Laboratory of Toxic Pathogens-Based Therapeutic Approaches of Gastric Cancer, Yangzhou 225000, China. Electronic address: taolide_2003@sina.com. 3. The Affiliated Hospital of Yangzhou University, Yangzhou University, Yangzhou 225000, China; The State Administration of Traditional Chinese Medicine Key Laboratory of Toxic Pathogens-Based Therapeutic Approaches of Gastric Cancer, Yangzhou 225000, China; Clinical Medicine College of Yangzhou University, Yangzhou 225000, China. Electronic address: yzuntyhybg@163.com. 4. The State Administration of Traditional Chinese Medicine Key Laboratory of Toxic Pathogens-Based Therapeutic Approaches of Gastric Cancer, Yangzhou 225000, China; Clinical Medicine College of Yangzhou University, Yangzhou 225000, China. Electronic address: guhao1980@126.com. 5. The Affiliated Hospital of Yangzhou University, Yangzhou University, Yangzhou 225000, China; The State Administration of Traditional Chinese Medicine Key Laboratory of Toxic Pathogens-Based Therapeutic Approaches of Gastric Cancer, Yangzhou 225000, China; Clinical Medicine College of Yangzhou University, Yangzhou 225000, China. Electronic address: jinfeng0522@126.com. 6. The State Administration of Traditional Chinese Medicine Key Laboratory of Toxic Pathogens-Based Therapeutic Approaches of Gastric Cancer, Yangzhou 225000, China; Clinical Medicine College of Yangzhou University, Yangzhou 225000, China. Electronic address: dxj2319@sina.com. 7. The Affiliated Hospital of Yangzhou University, Yangzhou University, Yangzhou 225000, China; The State Administration of Traditional Chinese Medicine Key Laboratory of Toxic Pathogens-Based Therapeutic Approaches of Gastric Cancer, Yangzhou 225000, China; Clinical Medicine College of Yangzhou University, Yangzhou 225000, China. Electronic address: 544968853@qq.com. 8. The Affiliated Hospital of Yangzhou University, Yangzhou University, Yangzhou 225000, China. Electronic address: chinadyb@vip.sina.com. 9. The Affiliated Hospital of Yangzhou University, Yangzhou University, Yangzhou 225000, China. Electronic address: wmxiao@yzu.edu.cn. 10. Department of Physiology, School of Medicine, Showa University, Tokyo 142, Japan. Electronic address: szgsy666@163.com. 11. Department of Physiology, School of Medicine, Showa University, Tokyo 142, Japan. Electronic address: 516545988@qq.com. 12. The State Administration of Traditional Chinese Medicine Key Laboratory of Toxic Pathogens-Based Therapeutic Approaches of Gastric Cancer, Yangzhou 225000, China; Clinical Medicine College of Yangzhou University, Yangzhou 225000, China. Electronic address: yyqian@yzu.edu.cn. 13. The Affiliated Hospital of Yangzhou University, Yangzhou University, Yangzhou 225000, China; The State Administration of Traditional Chinese Medicine Key Laboratory of Toxic Pathogens-Based Therapeutic Approaches of Gastric Cancer, Yangzhou 225000, China; Clinical Medicine College of Yangzhou University, Yangzhou 225000, China. Electronic address: yzumpi@163.com.
Abstract
ETHNOPHARMACOLOGICAL RELEVANCE: The traditional Chinese medicine (TCM) herb Celastrus orbiculatus is an important folk medicinal plant in China that has been used as an anti-inflammatory, antitumor, and analgesic in various diseases. The ethyl acetate extract of C. orbiculatus (C. orbiculatus extract, COE) was reported to show significant antitumor effects. However, no study in China or abroad has reported the effect and mechanism of COE in triggering apoptosis of gastric cancer (GC) cells. AIM OF STUDY: To further uncover the molecular mechanism underlying COE's apoptotic and anti-proliferative effects and lay a foundation for the development of novel, effective antitumor TCM agents. MATERIALS AND METHODS: The effect of COE on AGS and BGC-823 GC cell viability was examined using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Apoptosis of AGS and BGC-823 cells induced by COE was analyzed using flow cytometry and a mitochondrial membrane potential assay kit (JC-1). The proliferating GC cells were identified and examined using a 5-bromo-2'-deoxyuridine (BrdU) staining kit and flow cytometric analysis. A western blot assay was used to detect the effect of COE on apoptosis-related proteins, B-cell lymphoma-2 (Bcl-2), Bcl-extra-large (Bcl-xL), Bcl-2-like protein 12 (Bcl-L12), Bcl-2-associated X protein (Bax), and caspase as well as proliferation-related proteins, phosphoinositide 3-kinase (PI3K)/Akt/mechanistic target of rapamycin (mTOR)/p70s6k. Transmission electron microscopy (TEM) and an animal imaging technique were used to evaluate the microstructure of apoptotic GC cells and the effect of COE on tumor cell growth in vivo, respectively. RESULTS: The results indicate that COE significantly inhibited proliferation and induced apoptosis of GC AGS and BGC-823 cell lines both in vivo and in vitro. COE significantly decreased the cell mitochondrial membrane potential. Moreover, COE downregulated the levels of Bcl-2, Bcl-xL, and PI3K/Akt/mTOR/p70s6k while those of Bax and caspase were upregulated. More interestingly, COE altered the microstructure of the mitochondria. CONCLUSION: All these data collectively indicate that COE not only has significant antiproliferative effects but also has both in vivo and in vitro apoptotic effects. In addition, COE altered the structure and function of the mitochondria, which is another potential pathway for the antitumor activity of COE. These findings may provide a basis for the development of new anticancer TCM candidates.
ETHNOPHARMACOLOGICAL RELEVANCE: The traditional Chinese medicine (TCM) herb Celastrus orbiculatus is an important folk medicinal plant in China that has been used as an anti-inflammatory, antitumor, and analgesic in various diseases. The ethyl acetate extract of C. orbiculatus (C. orbiculatus extract, COE) was reported to show significant antitumor effects. However, no study in China or abroad has reported the effect and mechanism of COE in triggering apoptosis of gastric cancer (GC) cells. AIM OF STUDY: To further uncover the molecular mechanism underlying COE's apoptotic and anti-proliferative effects and lay a foundation for the development of novel, effective antitumor TCM agents. MATERIALS AND METHODS: The effect of COE on AGS and BGC-823 GC cell viability was examined using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Apoptosis of AGS and BGC-823 cells induced by COE was analyzed using flow cytometry and a mitochondrial membrane potential assay kit (JC-1). The proliferating GC cells were identified and examined using a 5-bromo-2'-deoxyuridine (BrdU) staining kit and flow cytometric analysis. A western blot assay was used to detect the effect of COE on apoptosis-related proteins, B-cell lymphoma-2 (Bcl-2), Bcl-extra-large (Bcl-xL), Bcl-2-like protein 12 (Bcl-L12), Bcl-2-associated X protein (Bax), and caspase as well as proliferation-related proteins, phosphoinositide 3-kinase (PI3K)/Akt/mechanistic target of rapamycin (mTOR)/p70s6k. Transmission electron microscopy (TEM) and an animal imaging technique were used to evaluate the microstructure of apoptotic GC cells and the effect of COE on tumor cell growth in vivo, respectively. RESULTS: The results indicate that COE significantly inhibited proliferation and induced apoptosis of GC AGS and BGC-823 cell lines both in vivo and in vitro. COE significantly decreased the cell mitochondrial membrane potential. Moreover, COE downregulated the levels of Bcl-2, Bcl-xL, and PI3K/Akt/mTOR/p70s6k while those of Bax and caspase were upregulated. More interestingly, COE altered the microstructure of the mitochondria. CONCLUSION: All these data collectively indicate that COE not only has significant antiproliferative effects but also has both in vivo and in vitro apoptotic effects. In addition, COE altered the structure and function of the mitochondria, which is another potential pathway for the antitumor activity of COE. These findings may provide a basis for the development of new anticancer TCM candidates.
Authors: Su Bin Park; Gwang Hun Park; Hun Min Song; Ho-Jun Son; Yurry Um; Hyun-Seok Kim; Jin Boo Jeong Journal: BMC Complement Altern Med Date: 2017-09-05 Impact factor: 3.659