Synthesizing oncogenic signal-processing systems that function as both "signal counters'' and "signal blockers'' in cancer cells
Synthesizing oncogenic signal-processing systems that function as both "signal counters'' and "signal blockers'' in cancer cells
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DOI：
10.1039/c3mb70093c
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发表时间：
2013-01-01
影响因子：
--
通讯作者：
Cai, Zhiming
中科院分区：
文献类型：
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作者：
Liu, Yuchen;Huang, Weiren;Cai, Zhiming
RNA-蛋白质相互作用在真核生物翻译调控中起着重要作用。这种现象提出了关于人工生物系统利用蛋白质-RNA相互作用的能力的问题。在这里，我们设计了一个致癌信号处理系统，表达海肾荧光素酶报告基因和萤火虫荧光素酶正常化基因，海肾荧光素酶报告基因在其50-非翻译区（5 '-UTR）中由RNA-蛋白质相互作用控制。为了测试所设计的系统的能力，我们随后构建了靶向核因子-κ B（NF-κ B）或β-连环蛋白信号的载体。我们发现荧光素酶表达的抑制（%）与靶蛋白含量相关，从而可以定量测量癌细胞中的致癌信号强度。该系统抑制了致癌信号下游基因的表达，并诱导膀胱癌细胞增殖抑制和凋亡，而不影响正常尿路上皮细胞。与传统方法（ELISA和定量免疫印迹）相比，生物系统提供了高度准确，一致和可重复的蛋白质信号定量，并能够区分癌细胞和非癌细胞。总之，合成系统在癌细胞中既起着“信号计数器”的作用，也起着“信号阻断剂”的作用。这种方法为致癌信号测量和癌症治疗提供了合成生物学平台。
RNA-protein interaction plays a significant role in regulating eukaryotic translation. This phenomenon raises questions about the ability of artificial biological systems to take the advantage of protein-RNA interaction. Here, we designed an oncogenic signal-processing system expressing both a Renilla luciferase reporter gene controlled by RNA-protein interaction in its 50-untranslated region (5'-UTR) and a Firefly luciferase normalization gene. To test the ability of the designed system, we then constructed vectors targeting the nuclear factor-kappa B (NF-kappa B) or the beta-catenin signal. We found that the inhibition (%) of luciferase expression was correlated to the targeted protein content, allowing quantitative measurement of oncogenic signal intensity in cancer cells. The systems inhibited the expression of oncogenic signal downstream genes and induced bladder cancer cell proliferation inhibition and apoptosis without affecting normal urothelial cells. Compared to traditional methods (ELISA and quantitative immunoblotting), the bio-systems provided highly accurate, consistent, and reproducible quantification of protein signals and were able to discriminate between cancerous and non-cancerous cells. In conclusion, the synthetic systems function as both "signal counters'' and "signal blockers'' in cancer cells. This approach provides a synthetic biology platform for oncogenic signal measurement and cancer treatment.