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Induction of Cytotoxicity and Apoptosis in Human Gastric Cancer Cell SGC-7901 by Isovaltrate Acetoxyhydrin Isolated from Patrinia heterophylla Bunge Involves a Mitochondrial Pathway and G2/M Phase Cell Cycle Arrest

  • Yang, Bo (College of Pharmaceutical Sciences, Zhejiang Chinese Medical University) ;
  • Wang, Yi-Qi (College of Pharmaceutical Sciences, Zhejiang Chinese Medical University) ;
  • Cheng, Ru-Bin (College of Pharmaceutical Sciences, Zhejiang Chinese Medical University) ;
  • Chen, Jia-Li (College of Pharmaceutical Sciences, Zhejiang Chinese Medical University) ;
  • Chen, Jin (Huadong Medicine Company Limited) ;
  • Jia, Li-Tao (Department of Gastroenterology, Tongde Hospital of Zhejiang Province) ;
  • Zhang, Ru-Song (College of Pharmaceutical Sciences, Zhejiang Chinese Medical University)
  • Published : 2013.11.30

Abstract

Background: Our previous study demonstrated cytotoxicity of a crude extract from Patrinia heterophylla Bunge (PHEB). In the present study, we aimed to investigate the effects of isovaltrate acetoxyhydrin (IA) isolated from PHEB on the gastric cancer cell SGC-7901, in order to explore a potential treatment for gastric cancer. Methods: MTT assays were employed to determine the effects of IA on cell vitality and proliferation, with monitoring of cell morphology changes and examination of apoptosis with Annexin V-PI staining. Flow cytometry was used to assess cell cycle progression and mitochondrial membrane potential. The activity of caspase 3, 9 was evaluated by spectrophotometry, and the protein levels of Bax, Bcl2 and Cyclin B1 were analyzed with Western blotting of total proteins extracted from cultured cells. Results: The results demonstrated direct toxicity of IA towards SGC-7901 cells. Evidence of apoptosis included blebbing and chromatin condensation. Annexin V-PI assays revealed early apoptosis, involving rapid depolarization of mitochondrial membranes and activity of caspase 3, 9 signaling pathways. Western blotting showed that Bcl2 and Bax proteins was down- and up-regulated, respectively, and cyclin B1 was up-regulated. Cell cycle analysis further indicated that IA could induce G2/M phase arrest in SGC-7901 cells. Conclusions: In conclusion, we believe that IA induces apoptosis of SGC-7901 cells, therefore providing a potential therapeutic agent for treatment of gastric cancer.

Keywords

Patrinia heterophylla Bunge;isovaltrate acetoxyhydrin;SGC-7901;apoptosis;G2/M arrest

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