عصاره آبی دانه گیاه آگرستما گیتاگو باعث مهار کسپاز 3 و توقف چرخه سلولی در فاز یک در رده سلولی AGS می شود.

Title

Aqueous Extract of Agrostemma githago Seed inhibits Caspase 3 and induces cell cycle arrest at G1 phase in AGS cell line.

English Abstract

ETHNOPHARMACOLOGICAL RELEVANCE Agrostemma githago is a weed plant belongs to Caryophyllaceae family which is used in folk medicine for treating cancers and warts. AIM OF THE STUDY This study was conducted to evaluate the cytotoxic effect of the aqueous extract of A. githago seed on gastric cancer cell line (AGS) and explore mechanism of apoptosis induction in the cells. MATERIALS AND METHODS A. githago seeds from suburb area of Ardabil Province, North West of Iran were collected and after aqueous extracting, dry matter harvested using lyophilizing technique. 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay was used for cytotoxicity test. Ethidium Bromide - Acridine Orange (EB/AO) staining method was used to detect apoptotic cells. Diamidino-2-phenylindole (DAPI) staining was used to analysis cell cycle using flow cytometer. AnnexinV binding level, Caspase 3 activity and Bcl-2 level were also measured to confirm apoptotic cell death. RESULTS After 24, 48 and 72 h of incubation with aqueous extract of A. githago seed, Cell growth was inhibited with IC50 values of 13.51 ± 0.7, 4.37 ± 1.01 and 2.42 ± 0.8 µg/ml, respectively. EB/AO staining method revealed that the extract exerts its cytotoxicity mainly via apoptosis which was in accordance with annexinV, blc-2 and caspase 3 results. The extract showed concentration depended increase in annexinV binding to externally exposed phosphatidylserine and also caspase 3 activity. The level of bcl-2 protein was decreased proportionally in respect to increasing level of the extract concentration. Cell cycle analysis revealed that the extract is able to arrest cells in G1 check point. CONCLUSION Our findings showed that aqueous extract of A. githago seed has cytotoxic effect on gastric cancer cells (AGS) mainly via apoptosis and froze cell cycle at G1 check point. It seems that the extract has promising effect on gastric cancer therapy in-vitro.

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