The study of Ziziphora Extract on Claudin1 Gene Expression in Cancerous Cell line AGS.

Claudins are the main structural and functional proteins of tight junctions in epithelial cells. The altered expression patterns of differ ent claudin members have been demonstrated in a variety of diseases, particularly in cancers.  It has been reported that claudin 1 is overexpressed 22-fold in gastric adenocarcinoma.  Considering the least side effect of herbal extracts, in this study we have evaluated  claudin-1 gene expression in AGS cells treated with ziziphora extract.

The AGS cells were incubated 37°C containing 5% CO2  with 85% humidity DMEM with  10% FBS .  The cells were treated with concentrations of 800, 1200, 2000 μg /ml of ziziphora for 48 and 72 hours. Extraction of RNA, synthesis of cDNA has been done using kit. The study of claudin-1 gene expression was performed by Real time PCR and also GAPDH gene was used as the internal control. Data analysis was performed with 2 - ΔΔCT and statistically analyzed by T-Test  .

The results of Real time PCR data have shown in 48 hours treatment, reduction of 7 and 2.3 fold with concentrations of 800 and 1200 μg /ml of ziziphora  respectively  and  in  72 hours treatment, reduction of 5.8 and 10 fold have seen in concentrations of 800 and 1200μg /ml of ziziphora, respectively and there was no significant  change in concentration of 2000μg / ml in both treatments.

Ziziphora extract in 1200 μg /ml concentration, reduced the expression of claudin-1 remarkably. Claudin-1 may become therapeutic targets for cancer treatment and ziziphora  as a herbal extract decreased the expression rate of claudin1significantly, in lower concentrations treatment.