State Key Laboratory of Oncology in South China, Collaborative Innovation Center for Cancer Medicine, Sun Yat-Sen University Cancer Center, Guangzhou 510060, P. R. China.
Markey Cancer Center, The University of Kentucky, College of Medicine, Lexington, Kentucky 40506, USA.
Theranostics. 2018 Apr 9;8(10):2739-2751. doi: 10.7150/thno.21477. eCollection 2018.
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. 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. Twist protein was proved to directly activate the transcription of 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. ROR1 is a targeted gene of Twist. Twist/ROR1 signaling is critical for invasion and metastasis of BLBC cells.
Twist 是诱导上皮-间质转化（EMT）的关键转录因子，促进细胞迁移、侵袭和癌症转移，赋予癌细胞干细胞样特征，并提供治疗抵抗。然而，Twist 在 EMT 和癌症进展中的功能作用和靶向基因仍然难以捉摸。通过微阵列分析从 T47D/Twist 细胞的全转录组中鉴定 Twist 的潜在靶向基因。通过 Western blot 和标记蛋白的免疫荧光检测 EMT 表型。采用双荧光素酶报告基因和染色质免疫沉淀检测实验观察 Twist 对 ROR1 的直接转录诱导作用。通过尾静脉注射 MDA-MB-231 细胞到裸鼠中，使用肺转移模型研究 Twist 和 ROR1 的促转移作用。生物信息学分析用于测量乳腺癌患者的无转移生存情况。 Twist 蛋白被证明可以直接激活非经典 WNT 信号通路中 Wnt5a 的受体基因的转录。沉默 ROR1 抑制基底样乳腺癌（BLBC）细胞的 EMT 过程、细胞迁移、侵袭和癌症转移。敲低 ROR1 也改善了 Twist 的促转移作用。此外，临床标本分析表明，ROR1 和 Twist 的高表达与乳腺癌患者无转移生存不良密切相关。 ROR1 是 Twist 的一个靶向基因。Twist/ROR1 信号通路对 BLBC 细胞的侵袭和转移至关重要。
