Down-regulation of DNA mismatch repair enhances initiation and growth of neuroblastoma and brain tumour multicellular spheroids.
Down-regulation of DNA mismatch repair enhances initiation and growth of neuroblastoma and brain tumour multicellular spheroids.
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DOI：
10.1371/journal.pone.0028123
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发表时间：
2011
期刊：
影响因子：
3.7
通讯作者：
Webb JS
中科院分区：
文献类型：
--
作者：
Collins SL;Hervé R;Keevil CW;Blaydes JP;Webb JS
多细胞肿瘤球体（MCTS）培养物是模拟体内肿瘤生长的发育和微环境条件的优秀模型系统。许多记录在案的细胞系在悬浮或非粘附环境中培养时可以产生分化的 MCTS。虽然 MCTS 的生理和生化特性已被广泛表征，但缺乏对导致这些结构启动的事件和条件的深入了解。 MCTS仅由一小部分细胞在表面相关生长过程中形成，但人们对这种分化的过程知之甚少，并且之前也没有进行过实验研究。对球体内基因表达的分析提供了线索，但迄今为止尚不清楚观察到的差异是 MCTS 生长的原因还是结果。与许多癌症中肿瘤发生相关的一种机制是 DNA 错配修复 (MMR) 受损引起的遗传不稳定性。本研究旨在确定 MMR 在 MCTS 启动和发展中的作用。使用表面相关的 N2a 和 CHLA-02-ATRT 培养系统，我们研究了 MMR 受损对 MCTS 生长的影响。对 DNA MMR 基因 MLH1 和 PMS2 的分析表明，与非球状体形成细胞相比，两者在 mRNA 水平上均显着下调。通过使用针对这些基因的小干扰 RNA (siRNA)，我们发现 MLH1 和 PMS2 的沉默增强了 MCTS 的启动和随后的扩展。 siRNA 转染后，这种效应在数代中得以延长。 DNA MMR 的下调可能导致 N2a 和 CHLA-02-ATRT MCTS 模型中肿瘤的发生和进展。表面相关 MCTS 分化的研究可能在研究癌灶发生过程中具有更广泛的应用。
Multicellular tumour spheroid (MCTS) cultures are excellent model systems for simulating the development and microenvironmental conditions of in vivo tumour growth. Many documented cell lines can generate differentiated MCTS when cultured in suspension or in a non-adhesive environment. While physiological and biochemical properties of MCTS have been extensively characterized, insight into the events and conditions responsible for initiation of these structures is lacking. MCTS are formed by only a small subpopulation of cells during surface-associated growth but the processes responsible for this differentiation are poorly understood and have not been previously studied experimentally. Analysis of gene expression within spheroids has provided clues but to date it is not known if the observed differences are a cause or consequence of MCTS growth. One mechanism linked to tumourigenesis in a number of cancers is genetic instability arising from impaired DNA mismatch repair (MMR). This study aimed to determine the role of MMR in MCTS initiation and development. Using surface-associated N2a and CHLA-02-ATRT culture systems we have investigated the impact of impaired MMR on MCTS growth. Analysis of the DNA MMR genes MLH1 and PMS2 revealed both to be significantly down-regulated at the mRNA level compared with non-spheroid-forming cells. By using small interfering RNA (siRNA) against these genes we show that silencing of MLH1 and PMS2 enhances both MCTS initiation and subsequent expansion. This effect was prolonged over several passages following siRNA transfection. Down-regulation of DNA MMR can contribute to tumour initiation and progression in N2a and CHLA-02-ATRT MCTS models. Studies of surface-associated MCTS differentiation may have broader applications in studying events in the initiation of cancer foci.
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影响因子：
--
作者：
Kerbel, R S
通讯作者：
Kerbel, R S
影响因子：
64.8
作者：
BRONNER, CE;BAKER, SM;LISKAY, RM
通讯作者：
LISKAY, RM
影响因子：
16
作者：
Coquelle, A;Toledo, F;Debatisse, M
通讯作者：
Debatisse, M
影响因子：
46.9
作者：
Helmlinger, G;Netti, PA;Jain, RK
通讯作者：
Jain, RK
影响因子：
13.8
作者：
Cahill, DP;Kinzler, KW;Lengauer, C
通讯作者：
Lengauer, C