Unravelling the metastasis-preventing effect of miR-200c in vitro and in vivo

Koehler, B.; Brieger, E.; Brandstaetter, T.; Hoerterer, E.; Wilk, U.; Poehmerer, J.; Joetten, A.; Paulitschke, P.; Broedersz, C. P.; Zahler, S.; Raedler, J. O.; Wagner, E.; Roidl, A.

doi:10.1101/2023.11.14.566527

bioRxiv · 10.1101/2023.11.14.566527

Unravelling the metastasis-preventing effect of miR-200c in vitro and in vivo

Koehler, B.·Brieger, E.·Brandstaetter, T.·Hoerterer, E.·Wilk, U.·Poehmerer, J.·Joetten, A.·Paulitschke, P.·Broedersz, C. P.·Zahler, S.·Raedler, J. O.·Wagner, E.·Roidl, A.

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Abstract

Advanced breast cancer as well as insufficient treatment can lead to the dissemination of malignant cells from the primary tumor to distant organs. Recent research has shown that miR-200c can hamper certain steps of the invasion-metastasis cascade. However, it is still unclear, whether sole miR-200c expression is sufficient to prevent breast cancer cells from metastasis formation. Hence, we performed a xenograft mouse experiment with inducible miR-200c expression in MDA-MB 231 cells. The ex vivo analysis of metastatic sites in a multitude of organs including lung, liver, brain, and spleen has revealed a dramatically reduced metastatic burden of mice with miR-200c expressing tumors. A fundamental prerequisite for metastasis formation is the motility of cancer cells and, therefore, their migration. Consequently, we analyzed the effect of miR-200c on collective and single cell migration in vitro, utilizing MDA-MB 231 and MCF7 cell systems with genetically modified miR-200c expression. Analysis of collective cell migration has resulted in confluence dependent motility of cells with altered miR-200c expression. Additionally, scratch assays have shown enhanced predisposition of miR-200c negative cells to leave cell clusters. The in-between stage of collective and single cell migration was validated using transwell assays, which have displayed reduced migration of miR-200c positive cells. Finally, to measure migration on single cell level, a novel assay on dumbbell shaped micropatterns was performed, which revealed that miR-200c critically determines confined cell motility. All of these results demonstrate that exclusive expression of miR-200c impedes metastasis formation in vivo and migration in vitro and highlight miR-200c as metastatic suppressor in breast cancer.

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Koehler, B., Brieger, E., Brandstaetter, T., Hoerterer, E., Wilk, U., Poehmerer, J., Joetten, A., Paulitschke, P., Broedersz, C. P., Zahler, S., Raedler, J. O., Wagner, E., Roidl, A.. 2023-11-16. Unravelling the metastasis-preventing effect of miR-200c in vitro and in vivo. https://doi.org/10.1101/2023.11.14.566527

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