The impact of octamer-binding transcription factor 4 and Nanog gene on pancreatic cancer stem cells in vivo

Abstract

Objective To studythe impact of octamer- binding transcription factor 4(Oct- 4) and Nanog gene on the biological characteristics of pancreatic cancer stem cells(PCSCs) in vivo. Methods The CD44+ CD24+ epithelial specific antigen(ESA)+ pancreatic cancer stem cells were sorted out via flow cytometry.The expression of Oct4 and Nanog genes in PCSCs were silenced by specific short hairpin RNA(shRNA)lentiviral vector.The interfering efficiency was detected by western blotting assay.The ectopic xenograft model was established via subcutaneous and intraperitoneal injection of Oct- 4 and Nanog silenced PCSCs, normal PCSCs and PANC- 1 cells into BALB/c nude mice, respectively. The influence of Oct4 and Nanog silencing on tumorigenicity, drug resistance and invasiveness were observed in vivo. Results The PCSCs accounted for 0. 1%-0. 8% in PANC- 1 cells.The efficiency of sh RNA silencing Oct4 and Nanog were(46. 00±0. 08)% and(78. 00±0. 12)% respectively.Silencing of Oct4 and Nanog significantly inhibited the tumorigenicity and invasiveness of PCSCs in nude mice and reversed the resistance to gemcitabine. Conclusion The lentiviral vector- mediated silencing of Oct4 and Nanog could inhibit the stem cell- like biological characteristics of PCSCs in vivo. Key words: Pancreatic cancer; Cancer stem cells; Octamer-binding transcription factor 4; Nanog; Gene silencing