Design and Functional Validation of a Mutant Variant of the LncRNA HOTAIR to Counteract Snail Function in Epithelial-to-Mesenchymal Transition.
Design and Functional Validation of a Mutant Variant of the LncRNA HOTAIR to Counteract Snail Function in Epithelial-to-Mesenchymal Transition.
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DOI：
10.1158/0008-5472.can-20-1764
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发表时间：
2021-01-01
期刊：
影响因子：
11.2
通讯作者：
Cicchini C
中科院分区：
文献类型：
--
作者：
Battistelli C;Garbo S;Riccioni V;Montaldo C;Santangelo L;Vandelli A;Strippoli R;Tartaglia GG;Tripodi M;Cicchini C
HOTAIR 是一种在多种上皮癌中过表达的 lncRNA，与侵袭密切相关。这种lncRNA被证明是上皮间质转化（EMT）的关键元件，EMT是一种触发转移的转分化过程。 Snail 是 EMT 的主要诱导剂，需要 HOTAIR 在特定上皮靶基因（即 HNF4α、E-钙粘蛋白和 HNF1α）上招募 EZH2 并导致其抑制。在这里，我们设计了一种 HOTAIR 缺失突变体形式，命名为 HOTAIR-sbid，包括假定的 Snail 结合域，但缺失了 EZH2 结合域。 HOTAIR-sbid 充当内源性 HOTAIR 的显性失活。在小鼠和人类肿瘤细胞中，HOTAIR-sbid 损害了 HOTAIR 结合 Snail 的能力，进而触发 H3K27me3/EZH2 介导的 Snail 上皮靶基因抑制。值得注意的是，HOTAIR-sbid 表达被证明可以降低细胞运动性、侵袭性、贴壁依赖性生长以及对 TGFβ 诱导的 EMT 的反应性。这些数据为基于 lncRNA 的策略提供了证据，该策略可有效损害主要 EMT 转录因子的功能。
HOTAIR is a lncRNA overexpressed in several epithelial cancers and strongly correlated with invasion. This lncRNA was proven a pivotal element of the epithelial-mesenchymal transition (EMT), a trans-differentiation process triggering metastasis. Snail, master inducer of EMT, requires HOTAIR to recruit EZH2 on specific epithelial target genes (i.e., HNF4α, E-cadherin and HNF1α) and cause their repression. Here we designed a HOTAIR deletion mutant form, named HOTAIR-sbid, including the putative Snail-binding domain but depleted of the EZH2 binding domain. HOTAIR-sbid acted as a dominant negative of the endogenous HOTAIR. In both murine and human tumor cells, HOTAIR-sbid impaired the ability of HOTAIR to bind Snail and, in turn, trigger H3K27me3/EZH2-mediated repression of Snail epithelial target genes. Notably, HOTAIR-sbid expression was proven to reduce cellular motility, invasiveness, anchorage-independent growth, and responsiveness to TGFβ-induced EMT. These data provide evidence on a lncRNA-based strategy to effectively impair the function of a master EMT-transcriptional factor.