Created Gli-1 duplex short-RNA (i-Gli-RNA) eliminates CD44 Hi progenitors of taxol-resistant ovarian cancer cells.-猫眼课题宝

DOI：

10.3892/or_00000793

发表时间：

2010-06

期刊：

Oncology reports

影响因子：

4.2

通讯作者：

中科院分区：

医学3区

文献类型：

作者：

Mine T;Matsueda S;Gao H;Li Y;Wong KK;Peoples GE;Ferrone S;Ioannides CG

来源链接：

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3426036/?report=classic

Notch和Hedgehog激活成体和癌症干细胞的细胞周期进程。Notch被DLL激活，Jag存在于相邻细胞上。我们研究了Notch激活配体Jag-1和靶向Gli-1的密度在紫杉醇/紫杉醇耐药（PTXRes）卵巢癌细胞SKOV 3（SKOV 3）分裂激活中的作用。我们使用特异性的γ-早老素抑制剂DAPT来鉴定Notch-1激活信号的特异性，并产生“蝶形双链体-3548-Gli-1-抑制性RNA”（i-Gli-1.RNA）来抑制细胞分裂。为了准确定量分裂动力学，在每个门控的分裂细胞群中测定CD 44和CD 24的表达。当被Jag-1 Low激活时，CD 44 High增殖，而当被Jag-1High激活时，CD 44 High增殖较差。DAPT抑制Jag-1 Low激活的细胞增殖，增加Jag-1High激活的细胞增殖。只有5-10%的细胞被Jag-1High和Jag-1 Low激活，分裂速度快，多项式，对称。i-Gli-1.RNA消除了第3次和第4次分裂中超过50%的小CD 44 High/CD 24 Neg细胞。与用阴性对照siRNA转染的细胞相比，该效应表现出特异性。i-Gli-1.RNA对大的CD 44 High/CD 24 Neg细胞没有影响，但抑制CD 44 High/CD 24 Low细胞的群体。活化自体肿瘤和纤维肉瘤细胞时，CD 44 High的扩增与Jag-1密度呈负相关。产生的i-RNA可能会减少静息CSC池。Notch和Gli-1信号在肿瘤干细胞的增殖/分裂和存活中起重要作用。通过其增强子G1 -1靶向Notch-1，对于消除紫杉醇抗性癌症干细胞（CSC）的新治疗应该是重要的。i.Gli-1 RNA如果与紫杉醇一起使用应该更有效。

Notch and Hedgehog activate cell-cycle progression of adult and cancer stem cells. Notch is activated by DLL and Jag presents on neighboring cells. We investigated the effects of density of the Notch-activating ligand, Jag-1, and targeting Gli-1, in activation of division of paclitaxel/taxol-resistant, (PTXRes) ovarian cancer cells SKOV3 (SKOV3). We used the specific γ-presenilin inhibitor, DAPT, to identify the specificity of activating signals for Notch-1 and created ‘butterfly-duplex-3548-Gli-1-inhibitory RNA’ (i-Gli-1.RNA) to inhibit cell division. To accurately quantify kinetics of division, the expression of CD44 and CD24 was determined in each gated population of divided cells. CD44High proliferated when activated by Jag-1Low and poorly when activated by Jag-1High. DAPT inhibited proliferation of cells activated by Jag-1Low, and increased proliferation of cells activated by Jag-1High. Only 5–10% of cells activated by Jag-1High and Jag-1Low divided fast, polynomial, and symmetric. i-Gli-1.RNA eliminated more than 50% of the small CD44High/CD24Neg cells in divisions 3 and 4. This effect appeared specific compared with cells transfected with negative control siRNA. i-Gli-1.RNA had no effect on large CD44High/CD24Neg cells, but inhibited the population of CD44High/CD24Low cells. Expansion of CD44High inversely correlated with Jag-1 density on activating autologous tumor and fibrosarcoma cells. Created i-RNAs may decrease the resting CSC pool. Notch and Gli-1 signals play an important role in proliferation/division and survival of cancer stem cells. Targeting Notch-1 through its enhancer Gl-1, should be significant for novel treatments to eliminate taxol-resistant cancer stem cells (CSC). i.Gli-1 RNA should be more effective if used together with Taxol.