Abstract
It has long been hypothesized that an increase in the extracellular matrix (ECM) stiffness mechanoactivates malignant phenotypes of breast tumor cells by regulating an array of processes underlying cancer biology. Although the contribution of substrate stiffening to drive malignant phenotype traits and other biological functions of a tumor is increasingly understood, the functional role of substrate softening on breast cancer cellular responses has rarely been investigated. Herein, we employed matrix metalloproteinase (MMP)-sensitive film to perform assays to explore the consequences of lowering stiffness on the biological behaviors of breast cancer cell MDA-MB-231. We demonstrated that cells underwent dramatic changes in migration, cellular conjunction, and expression of malignance-associated proteins and genes when the substrate stiffness decreased. Based on RNA sequencing and analysis, we found that hub genes including TP53, CCND1, MYC, CTNNB1, and YAP were validated to play central parts in regulating stiffness-dependent cellular manner change. Moreover, through visualization of differentially expressed genes (DEGs), cells on dynamically softened substrate appeared less influenced by transfer to tissue culture polystyrene (TCPS). These results suggest substrates with decreasing stiffness could normalize breast tumor malignant phenotype and help cells store the mechanical memory of the consequential weaker malignance.
摘要
肿瘤的发生发展伴随着细胞外基质的重塑和硬化. 目前, 尽 管细胞外基质的硬度已经成为一种强有力的肿瘤细胞行为调节器, 但基底硬度动态下降对乳腺癌细胞行为的影响却鲜有研究. 在此, 我们采用基质金属蛋白酶敏感的聚电解质多层膜进行分析, 探讨 硬度的降低对乳腺癌细胞MDA-MB-231生物学行为的影响. 结果 表明, 与在静态高硬度基底上培养的细胞相比, 在硬度动态降低的 基底上培养时, 细胞的迁移、粘附及恶性相关蛋白和基因表达水 平发生了显著变化, 肿瘤恶性降低. 根据蛋白质相互作用PPI网络 分析结果, 中枢节点基因TP53, CCND1, MYC, CTNNB1和YAP被 认为在硬度依赖性细胞行为变化中起着核心作用. 此外, 将在动态 软化基底上培养的细胞转移到高硬度材料上之后, 细胞仍保持较 弱的恶性, 表现出一定的记忆性.
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Acknowledgements
This work was supported by the National Natural Science Foundation of China (21875210), the National Key Research and Development Program of China (2016YFC1102203), the Natural Key Research and Development Project of Zhejiang Province (2018C03015), Zhejiang Provincial Ten Thousand Talents Program (2018R52001), the Fundamental Research Funds for the Central Universities (2020FZZX003-01-03), and the Higher Education Discipline Innovation Project (111 Project) (B16042).
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Ren KF and Ji J conceived and supervised the project; Hu M and Ren KF designed the experiments; Hu M and Li X carried out the experiments; Hu M wrote the manuscript. Ren KF revised the manuscript. All authors contributed to the general discussion.
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The authors declare that they have no conflict of interest.
Mi Hu is a PhD candidate in polymer material at Zhejiang University. Her research focuses on the construction of biocompatible polyelectrolyte films and hydrogels.
Xu Li is a PhD candidate in material engineering at Zhejiang University. His research focuses on the interaction between material and cells.
Ke-Feng Ren received his PhD degree from Zhejiang University in 2006. After working as a Postdoc with Prof. C. Picart in CNRS, France, he is now a full professor at Zhejiang University. He has co-authored over 100 peer-reviewed publications. His current research focuses on biomaterials interfaces, medical coating, cardiovascular medical devices.
Jian Ji received his PhD degree from Zhejiang University in 1997. In 2010, he received the Distinguished Young Scholars Award of the National Science Foundation of China. In 2015, he was awarded as Cheung Kong Scholars by the Ministry of Education. He has co-authored over 200 peer-reviewed publications. His research focuses on interfacial phenomena for biomedical implant, tissue engineering and nanomedicine.
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Hu, M., Li, X., Huang, WP. et al. Dynamically softened substrate regulates malignancy of breast tumor cells. Sci. China Mater. 64, 2580–2592 (2021). https://doi.org/10.1007/s40843-021-1631-3
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DOI: https://doi.org/10.1007/s40843-021-1631-3