Abstract
Insulin-like growth factor-1 (IGF-1) plays an important role in function and development of the mammary gland. However, high levels of IGF-1 has been associated with an increased risk of breast cancer development. Epithelial–mesenchymal transition (EMT) is a process where epithelial cells lose their epithelial characteristics and acquire a mesenchymal phenotype, which is considered one of the most important mechanisms in cancer initiation and promotion of metastasis. Extracellular vesicles (EVs) are released into the extracellular space by different cell types, which mediate intercellular communication and play an important role in different physiological and pathological processes, such as cancer. In this study, we demonstrate that EVs from MDA-MB-231 breast cancer cells stimulated with IGF-1 (IGF-1 EVs) decrease the levels of E-cadherin, increase the expression of vimentin and N-cadherin and stimulate the secretion of metalloproteinase-9 in mammary non-tumorigenic epithelial cells MCF10A. IGF-1 EVs also induce the expression of Snail1, Twist1 and Sip1, which are transcription factors involved in EMT. Moreover, IGF-1 EVs induce activation of ERK1/2, Akt1 and Akt2, migration and invasion. In summary, we demonstrate, for the first time, that IGF-1 EVs induce an EMT process in mammary non-tumorigenic epithelial cells MCF10A.
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Acknowledgements
We are grateful to Nora Ruiz and Maria de Lourdes-Rojas (LaNSE, Cinvestav-IPN) for their technical assistance in TEM.
Funding
This research was funded by CONACYT (255429) and CONACYT-FOSISS (Salud 2015-1-261637), Mexico. Grants from CONACYT supported E L-O, J R-R and AG-H.
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12079_2021_638_MOESM2_ESM.tif
IGF-1 Induces migration in MDA-MB-231 breast cancer cells. Migration assays were performed with MDA-MB-231 cells unstimuated (Basal) and stimulated with 1, 5, 10, 20 and 50 nM IGF-1 for 48. One control of migration was included (FBS). Graph represents the mean ± SD and indicate the fold of migration above basal value. *P˂0.05, ***P˂0.001. (TIF 1845 KB)
12079_2021_638_MOESM3_ESM.tif
Schematic diagram showing the general experimental procedure of stimulation of MDA-MB-231 cells with 10 nM IGF-1 for 12, 24, 36 and 48 h and the isolation of EV fractions, as well as the treatment of MCF10A or MDA-MB-231 cells with the EVs isolated. (TIF 1389 KB)
12079_2021_638_MOESM4_ESM.tif
Ctrl EVs do not regulate the expression of E- and N-cadherin. MCF10A cells were treated for various periods of time with Ctrl EVs and lysed. a and b Cell lysates were analyzed by WB with anti-E-cadherin Ab, anti-N-cadherin Ab and anti-actin Ab. One control of MCF10A cells without treatment with EVs was included (Basal). Graphs represent the mean ± SD and indicate the fold of E-cadherin and N-cadherin above basal value. ns: not significant. (TIF 498 KB)
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Leal-Orta, E., Ramirez-Ricardo, J., Garcia-Hernandez, A. et al. Extracellular vesicles from MDA-MB-231 breast cancer cells stimulated with insulin-like growth factor 1 mediate an epithelial–mesenchymal transition process in MCF10A mammary epithelial cells. J. Cell Commun. Signal. 16, 531–546 (2022). https://doi.org/10.1007/s12079-021-00638-y
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DOI: https://doi.org/10.1007/s12079-021-00638-y