بررسی اثرات الیگودزوکسی نوکلئوتیدها روی سلول های مدل سرطانی

Title

Potential Effects of CpG-Oligodeoxynucleotides on Intestinal Epithelial Cells

English Abstract

Objectives: A pair of synthetic un-methylated CpG ODNs were used to assess the potential modulatory effects on non immune cell (epithelial cell line HT-29). Material & Methods: epithelial cell line HT-29 was treated with CpG ODN(CpG 2006) and lipopolysaccharide (LPS) at (5, 10, 25, 50 µg/ml) and (1,5,10 µg/ml) concentrations, respectively .Following treatments, dose-response and time-course cytotoxicity (using a colorimetric method), MMP-2 activity (using gelatin zymography)and apoptosis (using annexin-v flowcytometry method)assays were performed. to verify specific CpG effects , Chloroquine treatment was used for its inhibitory effect on endosomal acidification process. Results: Cytotoxicity analysis of CpG –ODN showed that CpG –ODN increases proliferation of cells at concentration 10-25 µg/ml. significant differences were observed between treated and untreated groups(p<0.05). MMP-2 activity analysis showed no significant differences between different groups of treated and untreated cells. Moreover, rate of apoptosis for CpG –ODN treated cells at 5,10,25 µg/ml concentrations was 0.8%, 6.46% and 14.21% , respectively, showing an increased apoptosis to necrosis ratio. Conclusion: intestinal epithelial cell line HT-29 is potentially highly responsive to CpG therapy upon TLR9 ligand stimulation in vitro.

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