Differentiation-inducing effect of berberine on U251 glioma stem cells

Zou Yuhui; Wang Qiong; Wang Weimin

doi:10.11850/j.issn.1009-122X.2016.12.010

Abstract：Objective To investigate the effect of berberine on glioma stem cell (GSC) differentiation. Methods GSCs were sorted from U251 glioma cells through Magnetic Activated Cell Sorting (MACS) and cultured, the immunomagnetic beads were marked by CD133, and GSCs were identified by flow cytofluorometry. The inhibition of berberine on GSC proliferation was assessed with Cell Counting Kit-8, and GSCs were treated with berberine at different dosages (0.001, 0.01, 0.1 or 1μmol/L), then the cell optical density (OD) was detected. The expressions of stem cell markers (CD133, Nestin) and differentiated cell markers (GFAP, MAP2) were detected by real-time PCR (RT-PCR) and immunofluorescence. Results Over 98%sorted cells were CD133 positive cells after MACS, and the sorted cells could form neurospheres in serum-free medium. The Cell Counting Kit-8 test showed that the inhibition of berberine on the GSC growth was enhanced with the increase of berberine dosage and duration. Through RT-PCR test, it was found that berberine gradually decreased the expressions of CD133 and Nestin mRNA, and increased the expressions of GFAP and MAP2 mRNA with the time prolonged. Immunofluorescence assay showed that berberine depressed the expressions of CD133 and Nestin protein, and enhanced the expressions of GFAP and MAP2 protein. Conclusion Berberine can depress GSC proliferation effectively and induce its differentiation.