Resetting The Pancreatic Adenocarcinoma(PDAC) Cells into Non Tumorigenic Cells via Epigenetic ReprogrammingTechnology

Background: Although cancer is generally believed to developthrough accumulation of multiple genetic mutations, there is increasingevidence that cancer cells also acquire epigenetic abnormalitiesduring development, maintenance, and progression.By utilizing the reprogramming technology as a tool to introducethe ‘pressure’ to alter epigenetic regulations, we might beable to clarify the epigenetic behavior that is unique to cancercells.we hypothesized that using cell reprogramming technologywould allow the Pancreatic adenocarcinoma cells to exitfrom tumorigenicity.Materials and Methods: We therefore sought to reprogramPDX(patient derived xenograft) from human PDAC, by introducing(۱) lentiviral mediated induction of Yamanaka Factors(OSKM), (۲) the pluripotency associated gene OCT۴ and themicroRNA mir-۳۰۲ and (۳) Episomal vectors (OCT۴, SOX۲,KLF۴, LMYC and LIN۲۸A combined with P۵۳ knock-down(shP۵۳)) as a safe method to reprogram cells without genomeintegration. We compare these three different methods to findmost efficient and safe method to reprogram PDAC PDX cellsinto less or non-tumorigenic cells.Results: Immunostaining, Alkaline phosphates staining andreal time PCR showed that induction with the episomal vectorsis the most efficient method to reprogram our fibroblastand PDAC PDX cells and reprogramming of PDAC-PDX cellssignificantly altered their tumorigenic potential in vitro via differentiation-promoting effect of epigenetic reprogramming. Invivo results in nude mice clearly showed that direct reprogrammingdecreases the aggressiveness of PDAC-PDX ۲۴۷ cancercells as compared with its parental counterpart.Conclusion: This study demonstrated that the ReprogrammedPDAC PDX cancer cells were distinct from natural PDAC cellswith regard to their loss of tumorigenicity in vitro and in vivo.

کلیدواژه ها:

Epigenetic Reprogramming ، Pancreatic Adenocarcinoma ، Episomal Vectorts