Silencing CD۱۳۳ via siRNA reduces the ability of pancreatic cancer cells in stemness through OCT۴, SOX۲, Nanog genes regulation

Pancreatic cancer (PC) is one of the deadliest gastrointestinal cancers with very high metastasis and low survival rate (۱). Today, cancer stem cells (CSCs) have been proposed as one of the main causes of all types of cancers including (PC) (۲). In such a way that the resistance to common anti-cancer treatments such as chemotherapy and radiation and recurrence of resistant tumors is attributed to the high expression level of CD۱۳۳ as one of the specific markers on the surface of CSCs including PC and nuclear transcription factors such as OCT۴, SOX۲, Nanog as key role players in stem cell proliferation (۳). As an alternative treatment with high specificity to minimize side effects, silencing CD۱۳۳ gene by siRNA Suppression at the post-transcriptional level targeted to CSCs may be used for PC (۴). In the current study, we aimed to investigate the success rate of cytotoxic effect and reduction effect in stemness ability of PC cells and related genes expression followed by CD۱۳۳ siRNA Suppression on Mia-Paca cancer cell line. Methods: In the present investigation, initially, the Mia-Paca cell line was cultured, followed by the transfection of the specific siRNA targeting the CD۱۳۳ gene into cells. In the subsequent phase, utilizing a methylthiazole tetrazolium kit (MTT assay), the viability of the cells following siRNA-mediated downregulation of CD۱۳۳ was examined. Using an ELISA reader, the optical density (OD) of each well was determined at a wavelength of ۵۷۰ nm. Furthermore, the expression of OCT۴, SOX۲, Nanog, stem cell proliferation genes was evaluated in siRNA transfected cells by Real-time PCR following extraction of cDNA. The results were analyzed by Graphpad prism software. Result: Our results indicated that transfection ofCD۱۳۳ specific siRNA in Mia-Paca cells reduces the viability in transfected cells after ۴۸ hours of incubation compared to untransfected control group. Also the expression of OCT۴, SOX۲, Nanog was significantly reduced in transfected cells.Conclusion: CD۱۳۳-siRNA had significant effects on the induction of stemness ability of cells and related genes (SOX۲, OCT۴ and Nanog) in Mia-PaCa cells.

کلیدواژه ها:

Pancreatic cancer ، Stemness ، Gene therapy ، CD۱۳۳ ، siRNA