The basic helix-loop-helix transcription factor E47 reprograms human pancreatic cancer cells to a quiescent acinar state with reduced tumorigenic potential.
The basic helix-loop-helix transcription factor E47 reprograms human pancreatic cancer cells to a quiescent acinar state with reduced tumorigenic potential.
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DOI：
10.1097/mpa.0000000000000328
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发表时间：
2015-07
期刊：
影响因子：
2.9
通讯作者：
Itkin-Ansari P
中科院分区：
文献类型：
--
作者：
Kim S;Lahmy R;Riha C;Yang C;Jakubison BL;van Niekerk J;Staub C;Wu Y;Gates K;Dong DS;Konieczny SF;Itkin-Ansari P
文本中提供了补充的数字内容。胰腺导管腺癌(PDA)起源于静止的腺泡细胞，这些细胞获得Kras突变，失去碱性螺旋-环-螺旋(BHLH)转录因子的信号，经历腺泡-导管化生，并迅速获得增加的生长潜力。我们质疑PDA细胞是否可以通过改变关键转录程序来重新编程，使其恢复到原来静止的腺泡细胞状态。人类PDA细胞系被设计为表达一种可诱导形式的bHLH蛋白E47。通过基因芯片、定量聚合酶链式反应、免疫印迹、免疫组织化学、小干扰RNA、染色质免疫沉淀分析和小鼠细胞移植等方法研究基因的表达、生长和功能。在人类PDA细胞中，E47的活动触发稳定的G0/G1期停滞，这需要细胞周期蛋白依赖的激酶抑制剂p21和应激反应蛋白TP53INP1。同时，E47诱导腺泡消化酶的高水平表达，并前馈激活受bHLH因子MIST1调控的腺泡成熟网络。此外，在体外诱导人PDA细胞中的E47足以抑制肿瘤的形成。人的PDA细胞保持高度的可塑性，这可以被用来诱导一种静止的腺泡细胞状态，降低致瘤潜力。此外，bHLH活性是协调调节人PDA细胞生长与细胞命运的关键节点。
Supplemental digital content is available in the text. Pancreatic ductal adenocarcinoma (PDA) initiates from quiescent acinar cells that attain a Kras mutation, lose signaling from basic helix-loop-helix (bHLH) transcription factors, undergo acinar-ductal metaplasia, and rapidly acquire increased growth potential. We queried whether PDA cells can be reprogrammed to revert to their original quiescent acinar cell state by shifting key transcription programs. Human PDA cell lines were engineered to express an inducible form of the bHLH protein E47. Gene expression, growth, and functional studies were investigated using microarray, quantitative polymerase chain reaction, immunoblots, immunohistochemistry, small interfering RNA, chromatin immunoprecipitation analyses, and cell transplantation into mice. In human PDA cells, E47 activity triggers stable G0/G1 arrest, which requires the cyclin-dependent kinase inhibitor p21 and the stress response protein TP53INP1. Concurrently, E47 induces high level expression of acinar digestive enzymes and feed forward activation of the acinar maturation network regulated by the bHLH factor MIST1. Moreover, induction of E47 in human PDA cells in vitro is sufficient to inhibit tumorigenesis. Human PDA cells retain a high degree of plasticity, which can be exploited to induce a quiescent acinar cell state with reduced tumorigenic potential. Moreover, bHLH activity is a critical node coordinately regulating human PDA cell growth versus cell fate.