ATP citrate lyase knockdown impacts cancer stem cells in vitro.
ATP citrate lyase knockdown impacts cancer stem cells in vitro.
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DOI：
10.1038/cddis.2013.215
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发表时间：
2013-06-27
影响因子：
9
通讯作者：
中科院分区：
文献类型：
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ATP柠檬酸裂解酶（ACL）敲低（KD）导致肿瘤抑制并诱导分化。我们以前曾报道ACL KD逆转肺癌细胞的上皮间质转化（EMT）。由于EMT通常与诱导干细胞的过程相关，因此我们假设ACL KD影响癌症干细胞。通过评估肿瘤球的形成和干细胞标志物的表达，我们发现这是A549细胞的情况，其中含有Ras突变，以及其他两种非小细胞肺癌细胞系H1975和H1650，由激活EGFR突变驱动。诱导型ACL KD与稳定型ACL KD具有相同的效果。在另一个充分表征的Ras诱导的乳腺模型系统（HMLER）中观察到类似的作用。此外，用羟基柠檬酸盐处理表型模仿ACL KD的作用，表明ACL的酶活性是关键的。事实上，乙酸盐处理逆转了ACL KD表型。先前已经确定ACL KD通过磷脂酰肌醇3-激酶（PI 3 K）途径而不是Ras-有丝分裂原活化蛋白激酶（MAPK）途径影响信号传导，并且EMT可以被PI 3 K抑制剂逆转，我们惊讶地发现这些系统中的干性通过Ras-MAPK信号传导而不是通过PI 3 K信号传导维持。Snail是一种受Ras-MAPK信号传导影响的下游转录因子，已知可促进EMT和干性。我们发现，ACL KD降低了snail表达。在致瘤性HMLER细胞中，ACL过表达增加了snail表达和干性，这两者都被ACL KD降低。此外，ACL本身不能启动肿瘤发生或干性。ACL和snail蛋白相互作用，ACL表达调节snail的转录活性。最后，ACL KD抵消了由Ras-MAPK信号传导外的途径激活驱动的不同细胞系统中诱导的干细胞特征。我们的研究结果揭示了ACL功能的一个新方面，即其对广泛遗传多样性细胞类型中癌症干细胞性的影响。
ATP citrate lyase (ACL) knockdown (KD) causes tumor suppression and induces differentiation. We have previously reported that ACL KD reverses epithelial–mesenchymal transition (EMT) in lung cancer cells. Because EMT is often associated with processes that induce stemness, we hypothesized that ACL KD impacts cancer stem cells. By assessing tumorsphere formation and expression of stem cell markers, we showed this to be the case in A549 cells, which harbor a Ras mutation, and in two other non-small-cell lung cancer cell lines, H1975 and H1650, driven by activating EGFR mutations. Inducible ACL KD had the same effect as stable ACL KD. Similar effects were noted in another well-characterized Ras-induced mammary model system (HMLER). Moreover, treatment with hydroxycitrate phenocopied the effects of ACL KD, suggesting that the enzymatic activity of ACL was critical. Indeed, acetate treatment reversed the ACL KD phenotype. Having previously established that ACL KD impacts signaling through the phosphatidylinositol 3-kinase (PI3K) pathway, not the Ras-mitogen-activated protein kinase (MAPK) pathway, and that EMT can be reversed by PI3K inhibitors, we were surprised to find that stemness in these systems was maintained through Ras-MAPK signaling, and not via PI3K signaling. Snail is a downstream transcription factor impacted by Ras-MAPK signaling and known to promote EMT and stemness. We found that snail expression was reduced by ACL KD. In tumorigenic HMLER cells, ACL overexpression increased snail expression and stemness, both of which were reduced by ACL KD. Furthermore, ACL could not initiate either tumorigenesis or stemness by itself. ACL and snail proteins interacted and ACL expression regulated the transcriptional activity of snail. Finally, ACL KD counteracted stem cell characteristics induced in diverse cell systems driven by activation of pathways outside of Ras-MAPK signaling. Our findings unveil a novel aspect of ACL function, namely its impact on cancer stemness in a broad range of genetically diverse cell types.
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DOI：
10.1186/bcr920
发表时间：
2004
期刊：
Breast cancer research : BCR
影响因子：
--
作者：
Dontu G;Jackson KW;McNicholas E;Kawamura MJ;Abdallah WM;Wicha MS
通讯作者：
Wicha MS
影响因子：
10.5
作者：
Dontu, G;Abdallah, WM;Wicha, MS
通讯作者：
Wicha, MS
影响因子：
48
作者：
Lin, Tongxiang;Ambasudhan, Rajesh;Yuan, Xu;Li, Wenlin;Hilcove, Simon;Abujarour, Ramzey;Lin, Xiangyi;Hahm, Heung Sik;Hao, Ergeng;Hayek, Alberto;Ding, Sheng
通讯作者：
Ding, Sheng
影响因子：
7.4
作者：
Fillmore, Christine M.;Kuperwasser, Charlotte
通讯作者：
Kuperwasser, Charlotte
DOI：
10.1083/jcb.200203064
发表时间：
2002-08-05
期刊：
The Journal of cell biology
影响因子：
--
作者：
Hanai J;Gloy J;Karumanchi SA;Kale S;Tang J;Hu G;Chan B;Ramchandran R;Jha V;Sukhatme VP;Sokol S
通讯作者：
Sokol S