Initial Identification Of Prostate Cancer Stem-like Cell Through MACS

Objective: To sort and initially identify the CD133~+/CD44~+stem cellthrough MACS from human prostate cancer cell line PC-3cells.Methods: The expression difference of CD133and CD44on themembrane of PC-3cells was detected after MACS through flow cytometry(FCM); then the formation of spheres cultured by serum-free medium (SFM)was observed and the subsequent immunofluorescence (IF) assay wasperformed; we further distinguished the differences of the morphology、proliferation capacity of PC-3cell after MACS; detected the expression ofprostatic acid phosphatase (PAP) after the differentiation assay throughimmunohistochemical (IHC) and western blot.Results: The positive expressions of the CD133and CD44of the PC-3cell were (1.33±0.05)%and (0.87±0.06)%through FCM, while they were(84.82±0.07)%and (99.91±0.03)%respectively after MACS; the positiveexpressions of CD133and CD44remained after SFM cultured byimmunofluorescence (IF); the proliferation capacity of CD133~+/CD44~+cellwas higher than PC-3cell (t=11, P=0.008) and CD133~+/CD44~+cell after induced (t=40.1, P=0.001); the expression of the PAP of the CD133~+/CD44~+cell was positive after induced by TGF-β, while it was negative in theCD133~+/CD44~+cell not induced by TGF-β through IHC and Western blot.Conclusions: The CD133~+/CD44~+cell had some characteristics ofstem cell after MACS from PC-3cell line through initial functionalidentification, which could lay the foundation for the further functional andbiological research of prostate cancer stem cells.