The design and optimization of RNA trans-splicing molecules for skin cancer therapy
The design and optimization of RNA trans-splicing molecules for skin cancer therapy
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DOI：
10.1016/j.molonc.2013.08.005
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发表时间：
2013-12-01
影响因子：
6.6
通讯作者：
Bauer, Johann W.
中科院分区：
文献类型：
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作者：
Gruber, Christina;Koller, Ulrich;Bauer, Johann W.
通过RNA反式剪接靶向肿瘤标志物基因是诱导肿瘤细胞特异性死亡的一种有前途的手段。利用筛选系统，我们设计了RNA反式剪接分子（RTM）特异性结合SLCO 1B 3的前体mRNA，表皮大疱相关鳞状细胞癌（EB-SCC）的标记基因。特异性反式剪接导致SLCO 1B 3的内源性靶mRNA与RTM提供的自杀基因编码序列融合。使用表达SLCO 1B 3的小基因（SLCO 1B 3-MG）分析了含有HSV-tk的SLCO 183特异性RTMs在异源环境中的反式剪接潜力。通过半定量RT-PCR和Western blot分析检测嵌合SLCO 1B 3-tk的表达。细胞活力和凋亡测定证实RTMs诱导SLCO 1B 3-MG表达细胞中自杀基因介导的凋亡。先导RTM还显示其促进EB-SCC细胞中内源性SLCO 1B 3前mRNA的反式剪接反应的潜力，导致tk介导的细胞凋亡。我们假设通过我们的诱导性细胞死亡系统对RTM的预选择加速了能够诱导皮肤癌细胞中的肿瘤特异性细胞死亡的最佳RTM的设计。(C)2013年欧洲生物化学学会联合会。Elsevier B. V.出版，保留所有权利。
Targeting tumor marker genes by RNA trans-splicing is a promising means to induce tumor cell-specific death. Using a screening system we designed RNA trans-splicing molecules (RTM) specifically binding the pre-mRNA of SLCO1B3, a marker gene in epidermolysis bullosa associated squamous cell carcinoma (EB-SCC). Specific trans-splicing, results in the fusion of the endogenous target mRNA of SLCO1B3 and the coding sequence of the suicide gene, provided by the RTM. SLCO183-specific RTMs containing HSV-tk were analyzed regarding their trans-splicing potential in a heterologous context using a SLCO1B3 expressing minigene (SLCO1B3-MG). Expression of the chimeric SLCO1B3-tk was detected by semi-quantitative RT-PCR and Western blot analysis. Cell viability and apoptosis assays confirmed that the RTMs induced suicide gene-mediated apoptosis in SLCO1B3-MG expressing cells. The lead RTM also showed its potential to facilitate a trans-splicing reaction into the endogenous SLCO1B3 pre-mRNA in EB-SCC cells resulting in tk-mediated apoptosis. We assume that the pre-selection of RTMs by our inducible cell-death system accelerates the design of optimal RTMs capable to induce tumor specific cell death in skin cancer cells. (C) 2013 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.