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Mechanoresponsive patterns of KLF2, 4, 5, and 6 expression differ among subclones from a single mammary tumor
Rafaela Marocci Lima Pimenta1, Cara Skon-Hegg2, Teresa Rose-Hellekant1
1Department of Biomedical Sciences, University of Minnesota Medical School, Duluth, MN 55812, USA.
Abstract:
A number of Krüppel-like transcription factor (KLF) family members display mechanoresponsive behaviors, and function as mechanosensitive transcription factors. There are many normal and pathological conditions where their roles in mechanotransduction and mechanoadaptation are not well understood, however. In this study, two basic questions regarding KLF mechanoresponsiveness were addressed: 1) are KLF 2, 4, 5, and 6 expressed at different levels among subclones of tumor cells adapted to specific microenvironmental conditions; and 2) is the expression of these KLFs responsive to rapid changes in the physical environment? To address these questions, the heterogeneous and differentially metastatic murine mammary tumor subclones 4T1, 4T07, and 67NR were subjected to physical changes in their culture conditions, and KLF responses assessed. The results show that the expression of different KLFs exhibit distinct responses to reductions in cell tension, as well as cell detachment from 2D and 3D environments. KLF2 and 4 expression is rapidly and temporarily induced upon release of cells from a stiff 2D substrate into liquid suspension culture in all three subclones, and similar responses are observed in two of the subclones upon the release of tension in 3D collagen gel cultures. By contrast, expression patterns of KLF5 and 6 were generally less affected by physical changes in most, but not all, of the cell lines examined. These results support the concept that KLFs differentially participate in transducing physical differences among intratumoral neighborhoods into distinct responses among heterogeneous subclones, thereby contributing to tumor cell behavioral complexity.
Insights
Krüppel-like factors (KLFs) are mechanosensitive transcription factors. This study shows KLF2 and KLF4 expression rapidly changes with mechanical stress, unlike KLF5 and KLF6, impacting tumor cell complexity.
Area of Science:
- Cell biology
- Molecular biology
- Cancer research
Background:
- Krüppel-like factors (KLFs) are transcription factors known to be mechanosensitive.
- Their precise roles in mechanotransduction and mechanoadaptation within normal and pathological conditions remain unclear.
- Understanding KLF behavior is crucial for deciphering tumor cell adaptation and complexity.
Purpose of the Study:
- To investigate differential Krüppel-like factor (KLF) expression in tumor cell subclones under varying microenvironmental conditions.
- To determine if KLF expression responds to rapid alterations in the physical cellular environment.
- To elucidate the role of KLFs in mechanotransduction within heterogeneous tumor populations.
Main Methods:
- Utilized murine mammary tumor subclones (4T1, 4T07, 67NR) known for heterogeneity and differential metastasis.
- Manipulated physical culture conditions, including cell tension and substrate detachment in 2D and 3D environments.
- Assessed the expression levels of KLF2, KLF4, KLF5, and KLF6 in response to these physical changes.
Main Results:
- KLF2 and KLF4 expression showed rapid, transient induction upon release from stiff 2D substrates or reduction of tension in 3D collagen gels across all tested subclones.
- KLF5 and KLF6 expression patterns were generally less sensitive to physical environmental changes in most cell lines.
- Distinct KLF responses were observed correlating with reductions in cell tension and detachment from different substrates.
Conclusions:
- KLF family members exhibit differential responses to mechanical stimuli, indicating varied roles in mechanotransduction.
- KLFs likely contribute to tumor cell behavioral complexity by transducing physical microenvironmental cues into distinct subclone-specific responses.
- These findings highlight the importance of mechanical forces in shaping tumor heterogeneity and progression.
