The loss of apical-basal cell polarity is a necessary stage of the epithelial-mesenchymal transition (EMT). Polarized epithelial cells interact with the basement membrane (BM) and, in particular, with laminins, the major components of BM. Here, we examined the effect of the transition of colon cancer cells from 2D polarized state to non-polarized 3D state on the expression of EMT associated genes, as well as the role of laminins 332 and 411 (LM-332 and LM-411) in this process. The three studied cell lines, HT-29, HCT-116 and RKO, were found to have different sensitivity to cultivation conditions (2D to 3D changes) and to addition of laminins. One of the possible reasons for this may be a difference in the initial 2D state of the cells. In particular, it was shown that the cell lines were at different EMT stages. HT-29 exhibited more epithelial expression profile, RKO was more mesenchymal, and HCT-116 was in an intermediate state. The most laminin-sensitive cell line was HCT-116. The magnitude and the specificity of cell response to LM-332 and LM-411 depended on the expression pattern of laminins’ receptors. EMT gene expression profile was not substantially changed neither during the transition from 2D to 3D state, nor the presence of laminins’ isoforms. However, we detected changes in expression of SNAI1 and ZEB1 genes encoding transcription factors that control the EMT process. Notably, in all three studied cell lines, the expression of SNAI1 was enhanced in response to laminin treatment.
This is a preview of subscription content, access via your institution.
REFERENCES
Samatov T.R., Tonevitsky A.G., Schumacher U. 2013. Epithelial–mesenchymal transition: Focus on metastatic cascade, alternative splicing, non-coding RNAs and modulating compounds. Mol. Cancer. 12, 107.
Lamouille S., Xu J., Derynck R. 2014. Molecular mechanisms of epithelial–mesenchymal transition. Nat. Rev. Mol. Cell Biol. 15, 178–196.
Domogatskaya A., Rodin S., Tryggvason K. 2012. Functional diversity of laminins. Annu. Rev. Cell Dev. Biol. 28, 523–553.
Matlin K.S., Myllymäki S.-M., Manninen A. 2017. Laminins in epithelial cell polarization: Old questions in search of new answers. Cold Spring Harb. Perspect. Biol., a027920.
Marinkovich M.P. 2007. Tumour microenvironment: Laminin 332 in squamous-cell carcinoma. Nat. Rev. Cancer. 7, 370–380.
Yurchenco P.D. 2011. Basement membranes: Cell scaffoldings and signaling platforms. Cold Spring Harb. Perspect. Biol. 3, a004911–a004911.
Pastushenko I., Brisebarre A., Sifrim A., Fioramonti M., Revenco T., Boumahdi S., Van Keymeulen A., Brown D., Moers V., Lemaire S., De Clercq S., Minguijón E., Balsat C., Sokolow Y., Dubois C., et al. 2018. Identification of the tumour transition states occurring during EMT. Nature. 556, 463–468.
Maltseva D.V., Rodin S.A. 2018. Laminins in metastatic cancer. Mol. Biol. (Moscow). 52 (3), 350–371.
Doi M., Thyboll J., Kortesmaa J., Jansson K., Iivanainen A., Parvardeh M., Timpl R., Hedin U., Swedenborg J., Tryggvason K. 2002. Recombinant human laminin-10 (alpha5beta1gamma1): Production, purification, and migration-promoting activity on vascular endothelial cells. J. Biol. Chem. 277, 12741–12748.
Khaustova N.A., Maltseva D.V., Oliveira-Ferrer L., Stürken C., Milde-Langosch K., Makarova J.A., Rodin S., Schumacher U., Tonevitsky A.G. 2017. Selectin-independent adhesion during ovarian cancer metastasis. Biochimie. 142, 197–206.
Maltseva D.V., Krainova N.A., Khaustova N.A., Nikulin S. V., Tonevitskaya S.A., Poloznikov A.A. 2017. Biodistribution of viscumin after subcutaneous injection to mice and in vitro modeling of endoplasmic reticulum stress. Bull. Exp. Biol. Med. 163, 451–455.
Krainova N.A., Khaustova N.A., Makeeva D.S., Fedotov N.N., Gudim E.A., Ryabenko E.A., Shkurnikov M.U., Galatenko V.V., Sakharov D.A., Maltseva D.V. 2013. Evaluation of potential reference genes for qRT-PCR data normalization in HeLa cells. Appl. Biochem. Microbiol. 49, 743–749.
Maltseva D.V., Khaustova N.A., Fedotov N.N., Matveeva E.O., Lebedev A.E., Shkurnikov M.U., Galatenko V.V., Schumacher U., Tonevitsky A.G. 2013. High-throughput identification of reference genes for research and clinical RT-qPCR analysis of breast cancer samples. J. Clin. Bioinform. 3, 13.
Nieto M.A., Huang R.Y.-J., Jackson R.A., Thiery J.P. 2016. EMT: 2016. Cell. 166, 21–45.
Nikulin S.V, Raigorodskaya M.P., Poloznikov A.A., Zakharova G.S., Schumacher U., Wicklein D., Stürken C., Riecken K., Fomicheva K.A., Alekseev B.Y., Shkurnikov M.Y. 2018. Role of IGFBP6 protein in the regulation of epithelial-mesenchymal transition genes. Bull. Exp. Biol. Med. 164, 650–654.
Kudriaeva A., Galatenko V., Maltseva D., Khaustova N., Kuzina E., Tonevitsky A., Gabibov A., Belogurov A. 2017. The transcriptome of type I murine astrocytes under interferon-gamma exposure and remyelination stimulus. Molecules. 22, 808.
Barretina J., Caponigro G., Stransky N., Venkatesan K., Margolin A.A., Kim S., Wilson C.J., Lehár J., Kryukov G.V., Sonkin D., Reddy A., Liu M., Murray L., Berger M.F., Monahan J.E., et al. 2012. The Cancer Cell Line Encyclopedia enables predictive modelling of anticancer drug sensitivity. Nature. 483, 603–607.
Ramovs V., Te Molder L., Sonnenberg A. 2017. The opposing roles of laminin-binding integrins in cancer. Matrix Biol. 57–58, 213–243.
Thomson S., Petti F., Sujka-Kwok I., Mercado P., Bean J., Monaghan M., Seymour S.L., Argast G.M., Epstein D.M., Haley J.D. 2011. A systems view of epithelial–mesenchymal transition signaling states. Clin. Exp. Metastasis. 28, 137–155.
Qin Y., Rodin S., Simonson O.E., Hollande F. 2017. Laminins and cancer stem cells: Partners in crime? Semin. Cancer Biol. 45, 3–12.
Galatenko V.V., Maltseva D.V., Galatenko A.V., Rodin S., Tonevitsky A.G. 2018. Cumulative prognostic power of laminin genes in colorectal cancer. BMC Med. Genomics. 11, 9.
Rodin S.A., Maltseva D.V. 2017. Laminins in colorectal cancer: Expression, function, prognostic power and molecular mechanisms. Res. Pract. Med. J. 4, 73–78.
Author information
Authors and Affiliations
Corresponding author
Additional information
Translated by I. Shipounova
Abbreviations: BM, basement membrane; LM, laminin when the certain isoform is designated; EMT, epithelial-mesenchymal transition.
Rights and permissions
About this article
Cite this article
Maltseva, D.V., Makarova, J.A., Khristichenko, A.Y. et al. Epithelial to Mesenchymal Transition Marker in 2D and 3D Colon Cancer Cell Cultures in the Presence of Laminin 332 and 411. Mol Biol 53, 291–298 (2019). https://doi.org/10.1134/S0026893319020110
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1134/S0026893319020110