Jin Feng, Li Hong-sheng, Zhao Lei, Wei Yu-jia, Zhang Hao, Guo Yuan-jin, Pang Ran, Jiang Xiao-bing, Zhao Hong-yang
Department of Neurosurgery, Hospital Affiliated to Jining Medical College, Jining 272029, China.
Zhonghua Yi Xue Za Zhi. 2008 Aug 26;88(33):2312-6.
To isolate cancer stem cells glioblastoma cells and detect the expression of anti-apoptotic and multi-drug resistance-associated protein (MRP) genes thereof.
CD133 positive cells were isolated from human glioblastoma multiforme cells of the line TJ905 by immunomagnetic beads technique and nestin, beta-tubulin and GFAP expression were examined by Immunofluorescence staining. RT-PCR was used to detect the expression of livin, livinalpha, Livinbeta, Survivin, MRP1, and MRP3.
Only 0.21% of the TJ905 cells maintained in serum was CD133(+) and showed characteristics of cancer stem cells, positive in nestin. These cells maintained a sphere-like growth status in serum-free medium in vitro, and could self-renew, proliferate, conditionally differentiate into tubulin-beta(+) and GFAP(+) cells, and produce neurons as well as glial cells. The mRNA expression levels of livin, livinalpha, survivin, MRP1, and MRP3 of the TJ905 tumor stem cells were significantly lower than those f the of TJ905 cells.
Cancer stem cells can be isolated from TJ905 glioblastoma multiforme cells. However, the generating rate of the tumor stem cells is lower than that of the TJ905 cells, and the expression levels of anti-apoptotic and MRP genes are lower than those of the progenitor cells. Showing that cancer stem cells are not the solo factor to maintain tumor growth and resist apoptosis and to pump the anti-tumor drugs out of cells.
分离胶质母细胞瘤细胞中的癌干细胞，并检测其抗凋亡及多药耐药相关蛋白（MRP）基因的表达。
采用免疫磁珠技术从TJ905人多形性胶质母细胞瘤细胞系中分离CD133阳性细胞，通过免疫荧光染色检测巢蛋白、β-微管蛋白和胶质纤维酸性蛋白（GFAP）的表达。采用逆转录-聚合酶链反应（RT-PCR）检测生存素、生存素α、生存素β、Survivin、多药耐药相关蛋白1（MRP1）和多药耐药相关蛋白3（MRP3）的表达。
血清培养的TJ905细胞中仅0.21%为CD133(+)，具有癌干细胞特征，巢蛋白呈阳性。这些细胞在无血清培养基中体外维持球形生长状态，可自我更新、增殖，条件性分化为β-微管蛋白(+)和GFAP(+)细胞，并产生神经元和神经胶质细胞。TJ905肿瘤干细胞中生存素、生存素α、Survivin、MRP1和MRP3的mRNA表达水平显著低于TJ905细胞。
可从TJ905多形性胶质母细胞瘤细胞中分离出癌干细胞。然而，肿瘤干细胞的生成率低于TJ905细胞，其抗凋亡及MRP基因的表达水平低于祖细胞。表明癌干细胞并非维持肿瘤生长、抵抗凋亡及将抗肿瘤药物泵出细胞的唯一因素。
