New-generation taxoid SB-T-1214 inhibits stem cell-related gene expression in 3D cancer spheroids induced by purified colon tumor-initiating cells.
New-generation taxoid SB-T-1214 inhibits stem cell-related gene expression in 3D cancer spheroids induced by purified colon tumor-initiating cells.
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DOI：
10.1186/1476-4598-9-192
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发表时间：
2010-07-14
期刊：
影响因子：
37.3
通讯作者：
Ojima I
中科院分区：
文献类型：
--
作者：
Botchkina GI;Zuniga ES;Das M;Wang Y;Wang H;Zhu S;Savitt AG;Rowehl RA;Leyfman Y;Ju J;Shroyer K;Ojima I
越来越多的证据表明，大多数肿瘤是分级组织的，由一群启动肿瘤的肿瘤干细胞(CSCs)组成，负责肿瘤的发展、维持和耐药。以前我们已经证明，结肠癌细胞的CD133高/CD44高比例在功能、形态和基因组水平上都不同于它们的大体对应细胞。与大多数中等水平表达CD133、CD44和CD166的结肠癌细胞相比，高表达CD133和CD44的细胞具有多种干细胞特征，包括在体内和体外具有强大的自我更新能力，以及产生不同细胞表型的能力。本研究有两个目的：a)确定CD133高/CD44高的结肠癌细胞诱导的漂浮3D多细胞球体的干细胞相关基因组特征；b)评估新一代紫杉类化合物SB-T-1214诱导的CSC特异性改变。从三个独立的侵袭性结肠癌细胞系HCT116、HT29和DLD-1中分离出CSC表型。干细胞特异性聚合酶链式反应芯片分析(SABiosciences)显示，与普通细胞相比，纯化的CD133高/CD44高表达细胞诱导的结肠球中干细胞相关基因的表达显著上调。流式细胞仪分析表明，3D结肠球中含有一些少数细胞群，它们高水平表达Oct4、Sox2、Nanog和c-Myc，对于干细胞的多能性和自我更新是必不可少的。单用浓度为100nM~(-1)μM的SB-T-1214作用48h，不仅可诱导这三种结肠癌球体在3D培养中的生长抑制和细胞死亡，而且还可导致干细胞相关基因的大量抑制和多能基因表达的显著下调。用Western blotting验证了聚合酶链式反应阵列和流式细胞仪的数据。重要的是，在这种治疗方案中存活下来的活细胞不再能够诱导二次漂浮的球体，并表现出显著的形态异常。我们报道了新一代紫杉类化合物SB-T-1214对耐药致癌细胞CD133High/CD44High诱导的结肠癌球体具有显著的抗肿瘤活性，并有效地抑制了大多数干细胞相关基因的表达。我们的数据表明，以前观察到的SB-T-1214在体内对耐药结肠肿瘤的长期疗效可能是通过下调肿瘤细胞群体中多个干细胞相关基因的表达来解释的，此外，它还是一种已知的抗增殖癌细胞的有丝分裂毒药。
Growing evidence suggests that the majority of tumors are organized hierarchically, comprising a population of tumor-initiating, or cancer stem cells (CSCs) responsible for tumor development, maintenance and resistance to drugs. Previously we have shown that the CD133high/CD44high fraction of colon cancer cells is different from their bulk counterparts at the functional, morphological and genomic levels. In contrast to the majority of colon cancer cells expressing moderate levels of CD133, CD44 and CD166, cells with a high combined expression of CD133 and CD44 possessed several characteristic stem cell features, including profound self-renewal capacity in vivo and in vitro, and the ability to give rise to different cell phenotypes. The present study was undertaken for two aims: a) to determine stem cell-related genomic characteristics of floating 3D multicellular spheroids induced by CD133high/CD44high colon cancer cells; and b) to evaluate CSC-specific alterations induced by new-generation taxoid SB-T-1214. Selected CSC phenotype was isolated from three independent invasive colon cancer cell lines, HCT116, HT29 and DLD-1. A stem cell-specific PCR array assay (SABiosciences) revealed that colonospheres induced by purified CD133high/CD44high expressing cells display profound up-regulation of stem cell-related genes in comparison with their bulk counterparts. The FACS analysis has shown that the 3D colonospheres contained some minority cell populations with high levels of expression of Oct4, Sox2, Nanog and c-Myc, which are essential for stem cell pluripotency and self-renewal. Single administration of the SB-T-1214 at concentration 100 nM-1 μM for 48 hr not only induced growth inhibition and apoptotic cell death in these three types of colon cancer spheroids in 3D culture, but also mediated massive inhibition of the stem cell-related genes and significant down-regulation of the pluripotency gene expression. PCR array and FACS data were confirmed with western blotting. Importantly, viable cells that survived this treatment regimen were no longer able to induce secondary floating spheroids and exhibited significant morphological abnormalities. We report here that a new-generation taxoid SB-T-1214 possesses significant activity against colon cancer spheroids induced by and enriched with drug resistant tumorigenic CD133high/CD44high cells and efficiently inhibited expression of the majority of stem cell-related genes. Our data indicates that the previously observed long-term efficacy of SB-T-1214 against drug resistant colon tumors in vivo may be explained by the down-regulation of multiple stem cell-related genes in the tumorigenic cell population, in addition to its known efficacy as a mitotic poison against proliferating cancer cells.
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影响因子：
4.8
作者：
Frank, NY;Pendse, SS;Frank, MH
通讯作者：
Frank, MH
DOI：
10.1186/bcr2106
发表时间：
2008
期刊：
Breast cancer research : BCR
影响因子：
--
作者：
Grimshaw MJ;Cooper L;Papazisis K;Coleman JA;Bohnenkamp HR;Chiapero-Stanke L;Taylor-Papadimitriou J;Burchell JM
通讯作者：
Burchell JM
影响因子：
11.2
作者：
Frank, NY;Margaryan, A;Frank, MH
通讯作者：
Frank, MH
DOI：
10.1073/pnas.0703478104
发表时间：
2007-06-12
影响因子：
11.1
作者：
Dalerba, Piero;Dylla, Scott J.;Clarke, Michael F.
通讯作者：
Clarke, Michael F.
影响因子：
11.2
作者：
Collins, AT;Berry, PA;Maitland, NJ
通讯作者：
Maitland, NJ