Twist promotes tumor metastasis in basal-like breast cancer by transcriptionally upregulating ROR1.
Twist promotes tumor metastasis in basal-like breast cancer by transcriptionally upregulating ROR1.
复制标题
Twist 通过转录上调 ROR1 促进基底样乳腺癌的肿瘤转移
作者：
Cao J;Wang X;Dai T;Wu Y;Zhang M;Cao R;Zhang R;Wang G;Jiang R;Zhou BP;Shi J;Kang T
基本原理：Twist是诱导上皮-间质转化（EMT）的关键转录因子，其促进细胞迁移、侵袭和癌症转移，赋予癌细胞干细胞样特征，并提供治疗抗性。然而，Twist在EMT和癌症进展中的功能作用和靶基因仍然难以捉摸。方法：利用基因芯片技术从T47 D/Twist细胞的转录组中筛选Twist的潜在靶基因。EMT表型的检测采用蛋白质免疫印迹法和标记蛋白免疫荧光法。采用双荧光素酶报告基因和染色质免疫沉淀法观察Twist对ROR 1的直接转录诱导作用。通过将MDA-MB-231细胞注射到裸鼠的尾静脉中，使用肺转移模型来研究Twist和ROR 1的促转移作用。利用生物信息学分析来测量乳腺癌患者的无转移生存率。结果：Twist蛋白被证明直接激活非经典WNT信号通路中Wnt 5a受体ROR 1基因的转录。ROR 1基因沉默抑制基底样乳腺癌细胞的EMT过程、细胞迁移、侵袭和癌转移。ROR 1的敲低也改善了Twist的促转移作用。此外，对临床标本的分析表明，ROR 1和Twist的高表达与乳腺癌患者的低无转移生存率密切相关。结论：ROR 1是Twist的靶基因。Twist/ROR 1信号通路对BLBC细胞的侵袭和转移至关重要。
Rationale: Twist is a key transcription factor for induction of epithelial-mesenchymal transition (EMT), which promotes cell migration, invasion, and cancer metastasis, confers cancer cells with stem cell-like characteristics, and provides therapeutic resistance. However, the functional roles and targeted genes of Twist in EMT and cancer progression remain elusive. Methods: The potential targeted genes of Twist were identified from the global transcriptomes of T47D/Twist cells by microarray analysis. EMT phenotype was detected by western blotting and immunofluorescence of marker proteins. The dual-luciferase reporter and chromatin immunoprecipitation assays were employed to observe the direct transcriptional induction of ROR1 by Twist. A lung metastasis model was used to study the pro-metastatic role of Twist and ROR1 by injecting MDA-MB-231 cells into tail vein of nude mice. Bio-informatics analysis was utilized to measure the metastasis-free survival of breast cancer patients. Results: Twist protein was proved to directly activate the transcription of ROR1 gene, a receptor of Wnt5a in non-canonical WNT signaling pathway. Silencing of ROR1 inhibited EMT process, cell migration, invasion, and cancer metastasis of basal-like breast cancer (BLBC) cells. Knockdown of ROR1 also ameliorated the pro-metastatic effect of Twist. Furthermore, analyses of clinical specimens indicated that high expression of both ROR1 and Twist tightly correlates with poor metastasis-free survival of breast cancer patients. Conclusion: ROR1 is a targeted gene of Twist. Twist/ROR1 signaling is critical for invasion and metastasis of BLBC cells.