Combination E2F-1 and p53 gene transfer does not enhance growth inhibition in human squamous cell carcinoma of the head and neck.
Combination E2F-1 and p53 gene transfer does not enhance growth inhibition in human squamous cell carcinoma of the head and neck.
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发表时间：
1998-09
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通讯作者：
D. Frank;T. Liu;M. Frederick;G. Clayman
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作者：
D. Frank;T. Liu;M. Frederick;G. Clayman
现有大量数据认为野生型p53和E2F-1协同介导细胞凋亡，E2F-1介导的细胞凋亡在某些情况下是p53依赖性的，并且E2F-1可以诱导哺乳动物细胞中p53的积累。这些数据支持对野生型 p53 和 E2F-1 组合过度表达在人头颈鳞状细胞癌 (SCCHN) 中的生物学后果的研究，旨在开发诱导细胞凋亡的分子干预策略来治疗这种破坏性疾病。重组腺病毒（Ad）载体Ad-p53和Ad-E2F-1分别用于将野生型p53和E2F-1基因转移至SCCHN细胞系TU138和TU167中。用 p53、E2F-1 或两者转导的 SCCHN 细胞进行了体外生长分析，结果表明同时转 p53 和 E2F-1 基因不会导致生长抑制增强。为了解释基于 E2F-1 和 p53 之间潜在负分子相互作用的生长测定结果，进行了 Western 和 Northern 印迹分析，以研究下游 p53 反式激活基因 p21Waf1 和 BAX 在各种 p53 和 E2F-1 基因转移条件下的差异表达。 Western 免疫印迹表明 E2F-1 拮抗 p53 对 p21Waf1 和 BAX 的诱导，而 Northern 印迹表明这种干扰是翻译前调节的且依赖于 p53。免疫共沉淀测定证实野生型 p53 和 E2F-1 基因产物在我们的细胞系中形成蛋白质-蛋白质复合物。我们的体外数据表明，在 SCCHN 中，E2F-1 可能通过蛋白质-蛋白质复合物的形成来干扰 p53 反式激活基因的诱导。同时 p53 和 E2F-1 基因转移在 SCCHN 体外模型中没有治疗优势。
Ample data exist contending that wild-type p53 and E2F-1 cooperate to mediate apoptosis, that E2F-1-mediated apoptosis is p53 dependent in some situations, and that E2F-1 can induce accumulation of p53 in mammalian cells. These data support the investigation of the biological consequences of combined wild-typep53 and E2F-1 overexpression in human squamous cell carcinoma of the head and neck (SCCHN) for the purpose of developing apoptosis-inducing molecular intervention strategies for the management of this devastating disease. The recombinant adenovirus (Ad) vectors Ad-p53 and Ad-E2F-1 were used for wild-type p53 and E2F-1 gene transfers, respectively, into SCCHN cell lines TU138 and TU167. SCCHN cells transduced with either p53, E2F-1, or both underwent in vitro growth analysis, which revealed that simultaneous p53 and E2F-1 gene transfer did not result in enhanced growth inhibition. To explain our growth assay findings on the basis of potential negative molecular interactions between E2F-1 and p53, Western and Northern blotting analyses were performed to investigate the differential expression of the downstream p53-transactivated genes, p21Waf1 and BAX, under various p53 and E2F-1 gene transfer conditions. Whereas Western immunoblotting demonstrated that E2F-1 antagonized p53 induction of p21Waf1 and BAX, Northern blotting revealed that this interference was pretranslationally regulated and p53 dependent. Coimmunoprecipitation assay confirmed that the wild-type p53 and E2F-1 gene products formed protein-protein complexes in our cell lines. Our in vitro data demonstrated that in SCCHN, E2F-1 interferes with induction of p53-transactivated genes, probably through the formation of protein-protein complexes. Simultaneous p53 and E2F-1 gene transfer is not therapeutically advantageous in this in vitro model of SCCHN.