Department of Neurosurgery, Shanghai Tenth People's Hospital, Tongji University, No. 301 Middle Yanchang Road, Zabei District, Shanghai, 200072, China.
Cell Mol Neurobiol. 2013 Oct;33(7):943-51. doi: 10.1007/s10571-013-9960-5. Epub 2013 Jul 13.
GSPCs (glioma stem/progenitor cells) were isolated from U87 glioma cell lines by serum-free neural stem cell medium. Four concentrations (1, 2, 4, and 8 μmol/L) of ATRA (all-trans retinoic acid) were used to induce the differentiation of GSPCs in the medium with or without growth factors. The effect of ATRA on the differentiation of GSPCs was analyzed by flow cytometry, real-time-PCR, and immunofluorescence. The differentiation of GSPCs could be induced by 1 or 2 μmol/L ATRA when GSPCs were cultured in growth factor-free medium. The detection of real-time-PCR showed that the level of GFAP (glial fibrillary acidic protein) mRNA of differentiated GSPCs in the growth factor-free medium containing 1 μmol/L ATRA group was significantly higher than that in the control group, and there was no significant difference in the level of TUBB-3 mRNA between the two groups. The GSPCs suffered apoptosis in the growth factor-free medium containing 4 or 8 μmol/L ATRA. The differentiation of GSPCs could not be induced by ATRA when GSPCs were cultured in the medium containing growth factors. The percentage of cells in G0/G1 phase was 84.26 ± 2.24 %, and the percentage of apoptosis was 18.95 ± 2.53 % in experimental groups which was similar to those in the control group. In conclusion, ATRA has certain capacity to induce differentiation of GSPCs, while its effective concentration should be controlled strictly. The differentiation of GSPCs induced by ATRA cannot antagonize the formidable differential inhibition of epidermal growth factor and basic fibroblast growth factor.
GSPCs（神经胶质瘤干细胞/祖细胞）是从 U87 神经胶质瘤细胞系中分离出来的，采用无血清神经干细胞培养基。用四种浓度（1、2、4 和 8 μmol/L）的全反式维甲酸（all-trans retinoic acid，ATRA）在有或无生长因子的培养基中诱导 GSPCs 的分化。通过流式细胞术、实时 PCR 和免疫荧光分析 ATRA 对 GSPCs 分化的影响。当 GSPCs 在无生长因子的培养基中培养时，1 或 2 μmol/L 的 ATRA 可诱导 GSPCs 分化。实时 PCR 检测显示，1 μmol/L ATRA 存在于无生长因子的培养基中诱导分化的 GSPCs 的 GFAP（胶质纤维酸性蛋白）mRNA 水平明显高于对照组，两组之间 TUBB-3 mRNA 的水平无显著差异。在含有 4 或 8 μmol/L ATRA 的无生长因子培养基中，GSPCs 发生凋亡。当 GSPCs 在含有生长因子的培养基中培养时，ATRA 不能诱导 GSPCs 分化。实验组细胞处于 G0/G1 期的比例为 84.26 ± 2.24%，凋亡比例为 18.95 ± 2.53%，与对照组相似。总之，ATRA 具有一定的诱导 GSPCs 分化的能力，但应严格控制其有效浓度。ATRA 诱导的 GSPCs 分化不能拮抗表皮生长因子和碱性成纤维细胞生长因子的强大分化抑制作用。
