Silencing CDC۲۰ gene by siRNA-loaded nanoniosome in cancer cell therapy

Backgrounds: Small interfering RNA (siRNA) by targeting and degrading oncogenes orangiogenic genes may be able to inhibit tumor cell growth. Cell division cycle ۲۰ homolog(CDC۲۰) protein as a proto-oncogene binds to the anaphase-promoting complex/cyclosome(APC/C) and result in the promotion of chromosomal separation and mitosis process. Thisprotein is highly expressed in several human carcinomas. Encapsulation of CDC۲۰siRNA innanocarriers prevents its rapid degradation by serum nucleases and enhances its stability andsafety. The purpose of this study was the knockdown of CDC۲۰ expression by siRNA-loadednanoniosome as a novel treatment method.Materials and Methods: The human gastric cancer cell line, AGS was maintained in DMEMsupplemented with ۱۰% fetal bovine serum (FBS) and ۱۰۰ U/ml penicillin, ۱۰۰ μg/mlstreptomycin at ۳۷˚C and ۵% CO۲. CDC۲۰siRNA was also purchased from DharmaconResearch. At first, ۱×۱۰۴ cells/ml was plated on ۶-well plates.After reaching ۷۰% confluence, the cells were incubated with ۱۰۰ nM free CDC۲۰siRNAs and۱۰۰ nM nanonoisomal CDC۲۰siRNAs for ۷۲ h. Untreated cells were considered as control.Then, RNA extraction, cDNA synthesis, and real-time PCR analysis were done and results wereexpressed with ۲-ΔΔCt.Results: we found that CDC۲۰siRNA remarkably suppressed the expression of CDC۲۰ at themRNA level in AGS cells after ۷۲ h compared to control. Free CDC۲۰siRNA almost ۱۰% andCDC۲۰siRNA-loaded nanonoisome up to ۴۰% demonstrated the inhibition of CDC۲۰ expressionat the transcriptional level.Conclusion: It seems that the nanoniosomal CDC۲۰siRNA has a hopeful application for thetreatment of gastric cancer.

کلیدواژه ها:

siRNA ، Gastric cancer ، Niosome ، CDC۲۰