Nakai Eiichi, Park Kaechang, Yawata Toshio, Chihara Takahiro, Kumazawa Ayano, Nakabayashi Hiromichi, Shimizu Keiji
Department of Neurosurgery, Medical School, Kochi University, Kochi, Japan.
Cancer Invest. 2009 Nov;27(9):901-8. doi: 10.3109/07357900801946679.
We established a cancer stem (CS) cell line, U87CS, by means of spheroid culture of U87MG cells derived from glioblastoma (GBM) in neuronal stem cell medium. U87CS cells presented positive immunohistochemical staining for multidrug resistance (MDR)1 and CD133, a marker for a subset of leukemia and GBM CS cells. The gene expression of MDR1 and CD133 on U87CS cells increased by an average of 8.51 and 47.18 times, respectively, compared to the levels on U87MG cells by real-time quantitative RT-PCR. U87CS cells possessed stronger drug-resistance to conventional anti-cancer drugs, such as doxorubicin (Dox), etoposide (VP-16), carboplastin, and BCNU than U87MG cells. Double immunofluoresence staining showed co-expression of MDR1 and CD133 on U87CS cells transplanted into nude mice brains. In addition, we identified the crossreactivity of CD133 and MDR1 in a surgical specimen of GBM. Our results suggest that CS cells may be resistant to current chemotherapy and represent a novel target for GBM therapeutics.
我们通过在神经干细胞培养基中对源自胶质母细胞瘤（GBM）的U87MG细胞进行球状体培养，建立了一种癌症干细胞（CS）系U87CS。U87CS细胞对多药耐药（MDR）1和CD133呈现阳性免疫组化染色，CD133是白血病和GBM CS细胞亚群的标志物。通过实时定量逆转录聚合酶链反应（RT-PCR），与U87MG细胞上的水平相比，U87CS细胞上MDR1和CD133的基因表达分别平均增加了8.51倍和47.18倍。与U87MG细胞相比，U87CS细胞对常规抗癌药物，如阿霉素（Dox）、依托泊苷（VP-16）、卡铂和卡莫司汀（BCNU）具有更强的耐药性。双重免疫荧光染色显示移植到裸鼠脑内的U87CS细胞上MDR1和CD133共表达。此外，我们在GBM手术标本中鉴定了CD133和MDR1的交叉反应性。我们的结果表明，CS细胞可能对当前化疗耐药，并且是GBM治疗的新靶点。
