Cancer Center, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts, United States of America.
Department of Medicine, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts, United States of America.
PLoS Biol. 2018 Jun 20;16(6):e2004049. doi: 10.1371/journal.pbio.2004049. eCollection 2018 Jun.
The contribution of basal and luminal cells to cancer progression and metastasis is poorly understood. We report generation of reporter systems driven by either keratin-14 (K14) or keratin-8 (K8) promoter that not only express a fluorescent protein but also an inducible suicide gene. Transgenic mice express the reporter genes in the right cell compartments of mammary gland epithelia and respond to treatment with toxins. In addition, we engineered the reporters into 4T1 metastatic mouse tumor cell line and demonstrate that K14+ cells, but not K14- or K8+, are both highly invasive in three-dimensional (3D) culture and metastatic in vivo. Treatment of cells in culture, or tumors in mice, with reporter-targeting toxin inhibited both invasive behavior and metastasis in vivo. RNA sequencing (RNA-seq), secretome, and epigenome analysis of K14+ and K14- cells led to the identification of amphoterin-induced protein 2 (Amigo2) as a new cell invasion driver whose expression correlated with decreased relapse-free survival in patients with TP53 wild-type (WT) breast cancer.
基底细胞和腔细胞对癌症进展和转移的贡献还不甚清楚。我们报告了由角蛋白-14（K14）或角蛋白-8（K8）启动子驱动的报告系统的产生，该系统不仅表达荧光蛋白，还表达可诱导的自杀基因。转基因小鼠在乳腺上皮的正确细胞区室中表达报告基因，并对毒素治疗有反应。此外，我们将报告基因工程化到 4T1 转移性小鼠肿瘤细胞系中，并证明 K14+细胞，而不是 K14-或 K8+，在三维（3D）培养中均具有高度侵袭性，并且在体内具有转移性。用报告基因靶向毒素处理培养中的细胞或小鼠中的肿瘤，可抑制体内的侵袭行为和转移。对 K14+和 K14-细胞的 RNA 测序（RNA-seq）、分泌组和表观基因组分析导致鉴定出两性蛋白诱导蛋白 2（Amigo2）作为新的细胞侵袭驱动因子，其表达与 TP53 野生型（WT）乳腺癌患者无复发生存率降低相关。
