Santos Mafalda, Ferreira Marta, Oliveira Patrícia, Mendes Nuno, André Ana, Vieira André F, Nunes Joana B, Carvalho Joana, Rocha Sara, Azevedo Mafalda, Ferreira Daniel, Reis Inês, Vinagre João, Paredes Joana, Heravi-Moussavi Alireza, Lima Jorge, Máximo Valdemar, Burleigh Angela, Roskelley Calvin, Carneiro Fátima, Huntsman David, Oliveira Carla
I3S, Instituto de Investigação e Inovação em Saúde, 4200-135 Porto, Portugal.
Ipatimup, Instituto de Patologia e Imunologia Molecular da Universidade do Porto, 4200-465 Porto, Portugal.
Biology (Basel). 2022 Jul 12;11(7):1046. doi: 10.3390/biology11071046.
Transitions between epithelial and mesenchymal cellular states (EMT/MET) contribute to cancer progression. We hypothesize that EMT followed by MET promotes cell population heterogeneity, favouring tumour growth. We developed an EMT model by on and off exposure of epithelial EpH4 cells (E-cells) to TGFβ1 that mimics phenotypic EMT (M-cells) and MET. We aimed at understanding whether phenotypic MET is accompanied by molecular and functional reversion back to epithelia by using RNA sequencing, immunofluorescence (IF), proliferation, wound healing, focus formation and mamosphere formation assays as well as cell xenografts in nude mice. Phenotypic reverted epithelial cells (RE-cells) obtained after MET induction presented epithelial morphologies and proliferation rates resembling E cells. However, the RE transcriptomic profile and IF staining of epithelial and mesenchymal markers revealed a uniquely heterogeneous mixture of cell subpopulations with a high self-renewal ability. RE cell heterogeneity was stably maintained for long periods after TGFβ1 removal both in vitro and in large tumours derived from the nude mice. Overall, we show that phenotypic reverted epithelial cells (RE cells) do not return to the molecular and functional epithelial state and present mesenchymal features related to aggressiveness and cellular heterogeneity that favour tumour growth in vivo. This work strengthens epithelial cell reprogramming and cellular heterogeneity fostered by inflammatory cues as a tumour growth-promoting factor in vivo.
上皮细胞和间充质细胞状态之间的转变（EMT/MET）有助于癌症进展。我们假设EMT后接着MET会促进细胞群体异质性，有利于肿瘤生长。我们通过将上皮EpH4细胞（E细胞）间歇性暴露于TGFβ1来建立一个EMT模型，该模型模拟了表型EMT（M细胞）和MET。我们旨在通过RNA测序、免疫荧光（IF）、增殖、伤口愈合、集落形成和乳腺球形成试验以及裸鼠体内细胞异种移植，了解表型MET是否伴随着分子和功能上恢复为上皮细胞状态。MET诱导后获得的表型恢复的上皮细胞（RE细胞）呈现出类似于E细胞的上皮形态和增殖率。然而，RE细胞的转录组图谱以及上皮和间充质标志物的IF染色显示，存在具有高自我更新能力的独特异质细胞亚群混合物。在去除TGFβ1后，RE细胞的异质性在体外和源自裸鼠的大肿瘤中都能长期稳定维持。总体而言，我们表明表型恢复的上皮细胞（RE细胞）不会恢复到分子和功能上皮状态，而是呈现出与侵袭性和细胞异质性相关的间充质特征，这有利于体内肿瘤生长。这项工作强化了炎症信号作为体内肿瘤生长促进因子所促进的上皮细胞重编程和细胞异质性。
