title: Aqueous extract of Agrostemma githago seed inhibits caspase-3 and induces cell-cycle arrest at G1 phase in AGS cell line.
abstract: BACKGROUND
  A weed plant belonging to the Caryophyllaceae family, Agrostemma githago is used in folk medicine to treat cancers and warts.
  
  
  OBJECTIVE
  The aim of this study was to evaluate the cytotoxic effect of the aqueous extract of A. githago seed on gastric cancer cell line (AGS) and to investigate the mechanism of apoptosis induction in these cells.
  
  
  METHODS
  Seeds of A. githago were collected from the suburban area of Ardabil Province, northwest Iran. After preparing the aqueous extract, dry matter was harvested with the lyophilizing technique. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to assess the cytotoxicity. Apoptotic cells were detected by staining with ethidium bromide/acridine orange (EB/AO). Diamidino-2-phenylindole (DAPI) staining was used for cell-cycle analysis with a flow cytometer. The annexin V binding level, caspase-3 activity, and B-cell lymphoma-2 (Bcl-2) level were also measured to confirm apoptotic cell death.
  
  
  RESULTS
  After the aqueous extract of A. githago seed was incubated for 24, 48, and 72 h, inhibited cell growth was observed with IC50 values of 13.51 ± 0.7, 4.37 ± 1.01, and 2.42 ± 0.8 µg/ml, respectively. The EB/AO staining method demonstrated that the extract exerts its cytotoxic effect mainly via apoptosis, in accordance with the annexin V, blc-2, and caspase-3 results. The extract showed a concentration-dependent increase in annexin V binding to externally exposed phosphatidylserine as well as caspase-3 activity. The bcl-2 protein level showed a proportionate decrease with the increase in extract concentration. The cell-cycle analysis revealed that the extract can arrest cells at the G1 checkpoint.
  
  
  CONCLUSIONS
  The findings of this study revealed the cytotoxic effect of the aqueous extract of A. githago seed on gastric cancer cells (AGS) mainly via apoptosis and the cell-cycle arrest at the G1 checkpoint. Therefore, the extract can be potentially used in gastric cancer therapy in vitro.
authors.affiliation: Department of Pharmacology and Toxicology, School of Pharmacy, Ardabil University of Medical Sciences, Ardabil, Iran; Drug and Advanced Sciences Research Center, School of Pharmacy, Ardabil University of Medical Sciences, Ardabil, Iran. Electronic address: shahab.bohlooli@arums.ac.ir.
authors.forename: Shahab
authors.initials: S
authors.lastname: Bohlooli
authors.affiliation: Department of Pharmacology and Toxicology, School of Pharmacy, Ardabil University of Medical Sciences, Ardabil, Iran.
authors.forename: Shahriar
authors.initials: S
authors.lastname: Bohlooli
authors.affiliation: Department of Pharmacology and Toxicology, School of Pharmacy, Ardabil University of Medical Sciences, Ardabil, Iran; Drug and Advanced Sciences Research Center, School of Pharmacy, Ardabil University of Medical Sciences, Ardabil, Iran; School of Agriculture, University of Mohaghegh Ardabili, Ardabil, Iran.
authors.forename: Roghaye
authors.initials: R
authors.lastname: Aslanian
authors.affiliation: Department of Pharmacology and Toxicology, School of Pharmacy, Ardabil University of Medical Sciences, Ardabil, Iran.
authors.forename: Fatemeh
authors.initials: F
authors.lastname: Nouri
authors.affiliation: School of Agriculture, University of Mohaghegh Ardabili, Ardabil, Iran.
authors.forename: Ali
authors.initials: A
authors.lastname: Teimourzadeh
mesh_terms: D031242:Agrostemma; D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D053148:Caspase 3 / Q000378:metabolism; D059447:Cell Cycle Checkpoints / Q000187:drug effects; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D016193:G1 Phase / Q000187:drug effects; D006801:Humans; D010936:Plant Extracts / Q000494:pharmacology*; D012639:Seeds
chemical_list: D000970:Antineoplastic Agents; D010936:Plant Extracts; D053148:Caspase 3
keywords: AGS cell line; Agrostemma githago; Cytotoxicity; Gastric cancer