CD133是一種胞外穿膜的蛋白質(或稱promonin-1)會表現在許多不同組織幹細胞，其中包含了腦腫瘤幹細胞。然而，CD133在腫瘤細胞的生理影響仍未清楚地了解。本實驗室利用慢病毒基因傳輸系統轉殖人類的CD133基因到大鼠膠質瘤細胞株C6 (hCD133+-C6)，作為研究癌化的模式細胞；先前的研究實驗顯示，表現人類CD133基因的C6細胞，其增殖及遷移能力有顯著性增加，並增強腫瘤的生成。本實驗進一步發現在C6細胞中過度表現人類CD133基因，會顯著性的增加其在軟膠中形成細胞群落的能力，並且增加活化的Notch1蛋白表現和傳遞Notch訊息的Hes1基因表現，而這樣的表現量增加可能對於hCD133+-C6增加的細胞增殖速度扮演一個重要的角色。而處理三氧化二砷(As2O3)，一種用於急性骨髓性白血病以及其他腫瘤的藥物，會導致hCD133+-C6隨著處理濃度、處理時間而降低其細胞存活率，並也發現三氧化二砷的處理能使其Hes1基因表現和Notch1的活化下降，然而三氧化二砷並不會顯著地影響C6-Puro的細胞存活率、Hes1 基因表現和活化的Notch1蛋白表現。進一步以軟膠細胞群落形成分析，發現三氧化二砷的前處理能降低hCD133+-C6在軟膠中形成細胞群落的能力。在動物實驗中，也發現三氧化二砷的前處理能降低hCD133+-C6在成鼠腦中腦腫瘤形成的體積。以上結果顯示，穩定表現人類CD133後，C6神經膠質瘤細胞對於三氧化二砷所誘發細胞死亡的敏感性增加，並且三氧化二砷可以抑制其腫瘤形成能力和Notch1/Hes1的表現。這樣的結果顯示Notch1/Hes1可能參與在hCD133+-C6的細胞增殖與細胞存活，而Notch1/Hes1可望作為一項治療具CD133表現神經膠質瘤細胞的可能標的。

CD133 (Prominin-1) is known as a cell surface marker found on multipotent stem cells, as well as brain tumor stem cells (tumor SCs). Through stable transduction by human CD133 using lentiviral gene delivery method, we have generated genetic modified C6 glioma cell line (hCD133+-C6). Previous studies have found overexpression of hCD133 increased cell proliferation, and migration. Moreover, implantation of hCD133+-C6 into rat cortex improved tumor formation. Herein by soft agar colony formation, we found that overexpression of hCD133 increased cell colony formation when compared to that observed C6 cells lacking hCD133 (C6-Puro). We describe that overexpression of hCD133 caused an increase in the levels of active Notch1 protein and in the gene expression of Hairy-enhancer of split1 (Hes1), a Notch effector. This increase may play a critical regulator for increased cell proliferation of hCD133+-C6 cells. In addition, exposure to arsenic trioxide (As2O3), a compound used for cure of acute promyelocytic leukemia and many solid tumors, dose and time dependently reduced the cell viability of hCD133+-C6 cells. The reduction was also associated with the downregulation of Hes1 mRNA expression and Notch1 activation. However, cell viability, Hes1 mRNA expression and active Notch1 levels in C6-Puro were not affected by arsenic trioxide. The pretreatment of arsenic trioxide on hCD133+-C6 also reduced the cell colony formation and tumor size in rat cortex. The results indicate that Notch1/Hes1 signaling is involved in the cell expansion and survival of hCD133+-C6 cells, suggesting that Notch1/Hes1 signaling can be a potential target for the treatment of CD133+-glioma cells.

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