Sun Yaxuan, Zhou Xun, Hu Xiaohui
Department of Pathology, School of Basic Medical Sciences, Anhui Medical University, Hefei 230032, China.
Department of Pathophysiology, School of Basic Medical Sciences, Anhui Medical University, Hefei 230032, China.
Biol Open. 2024 Dec 15;13(12). doi: 10.1242/bio.061790. Epub 2024 Dec 9.
Epithelial to mesenchymal transition (EMT) has been shown to play an essential role in the early stages of cancer cell invasion and metastasis. Inducible EMT models can initiate EMT in a controlled manner, thereby providing the opportunity to determine whether a cancer-associated gene influences cancer metastasis by triggering EMT. Moreover, different inducible EMT models enable the investigation of specific mechanisms of EMT modulation by various genes, facilitating a more precise understanding of how these genes influence cancer metastasis through the induction of EMT. Unfortunately, current inducible EMT models still present unmet needs. Therefore, we aimed to establish an inducible EMT model in MCF10A cells, a spontaneously immortalized human fibrocystic mammary cell line, by manipulating the expression of mouse Twist1 (mTwist1). In this study, we first compared the EMT induction capacity between human TWIST1 (hTWIST1) and mTwist1, and selected mTwist1 for further investigation. By monitoring the changes in epithelial and mesenchymal markers at different induction time points, we examined the EMT process in both polyclonal and monoclonal MCF10A cells that express doxycycline (DOX)-inducible mTwist1. Furthermore, our results showed that doxycycline-induced mTwist1 expression triggered EMT at a similar rate to TGFβ1-induced EMT in MCF10A cells. Additionally, this process was reversible upon DOX withdrawal. Thus, we have established a robust inducible EMT model in MCF10A cells, which can be used to further study cancer metastasis-driving genes.
上皮-间质转化（EMT）已被证明在癌细胞侵袭和转移的早期阶段起着至关重要的作用。可诱导的EMT模型能够以可控的方式启动EMT，从而提供机会来确定一个癌症相关基因是否通过触发EMT影响癌症转移。此外，不同的可诱导EMT模型能够研究各种基因对EMT调节的具体机制，有助于更精确地理解这些基因如何通过诱导EMT影响癌症转移。不幸的是，目前的可诱导EMT模型仍存在未满足的需求。因此，我们旨在通过操纵小鼠Twist1（mTwist1）的表达，在MCF10A细胞（一种自发永生化的人纤维囊性乳腺细胞系）中建立一个可诱导的EMT模型。在本研究中，我们首先比较了人TWIST1（hTWIST1）和mTwist1之间的EMT诱导能力，并选择mTwist1进行进一步研究。通过监测不同诱导时间点上皮和间质标志物的变化，我们检测了表达强力霉素（DOX）诱导型mTwist1的多克隆和单克隆MCF10A细胞中的EMT过程。此外，我们的结果表明，强力霉素诱导的mTwist1表达在MCF10A细胞中触发EMT的速率与TGFβ1诱导的EMT相似。此外，在撤除DOX后，这个过程是可逆的。因此，我们在MCF10A细胞中建立了一个强大的可诱导EMT模型，可用于进一步研究癌症转移驱动基因。
