Effect of intervention of Hedgehog signaling pathway on proliferation of human glioma stem cells in vitro
GONG Song-feng
DING Jian-jun
WANG Yu
SU Juan
WANG Xin-jun
WANG Mao-ming
LI Wei-ping
GAO Yong-zhong
Abstract：Objective To investigate the expression of Hedgehog signaling pathway in glioma stem cells (GSCs) and the effects of cyclopamine (a inhibitor of Hedgehog signaling pathway) on the proliferation of GSCs. Methods The GSCs were isolated and cultured from the glioblastomas removed by neurosurgery in our hospital. Immunohistochemical technique and fluorescence stain were used to identify GSCs and detect the expression of Hedgehog signaling pathway in GSCs. The effects of cyclopamine on the proliferation of GSCs were analysized by CCK-8. Results The suspended cell masses cultured by serum-free technique expressed CD33 and Nestin cell surface markers with strong proliferative and self-renewing ability, and was able to differentiated into glioma cells by the induction of 10%fetal bovine serum medium. Sonic Hedgehog and Smo, key proteins of Hedgehog signaling pathway, were expressed in GSCs. The results of CCK-8 showed that the inhibitory rates of 0.5, 1 and 5μmol/L cyclopamine to GSCs were (21.93±6.57)%, (32.03±9.13)%and (7.78±16.09)%respectively (P<0.01). The inhibitory rate of cyclopamine to GSCs 4, 12, 24, 36, 48 and 60 hours after the culture in the media contenting cyclopamine were (25.89±7.11)%, (28.00±10.00)%, (32.98±16.87)%, (35.83±17.44)%and (41.31±16.12)%, (41.58± 19.72)% respectively (P<0.01). Conclusion Cyclopamine, the inhibitor of Hedgehog signaling pathway, can significantly inhibits the proliferation of GSCs in the concentration-and time-depenent fashions.
Keywords：Glioma stem cellHedgehog signaling pathwayCyclopamineProliferation
Publication Date：2015-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：3( 96-98 )