INDUCTION OF EPITHELIAL-TO-MESENCHYMAL TRANSITION IN MCF-7-SNAI1 CELLS LEADS TO REORGANIZATION OF ADHERENS JUNCTIONS AND ACQUISITION OF MIGRATORY ACTIVITY
https://doi.org/10.21294/1814-4861-2018-17-4-24-29
Abstract
Using DIC and confocal microscopy, changes in morphology, migratory characteristics and adherence junctions (AJs) were analyzed in the mammary carcinoma cell line MCF-7-SNAI1 after activation of the EMT transcription factor SNAI1. Western Blot analysis showed that after removal of tetracycline from the cell culture medium expression of SNAI1 reached its peak in 24 hours and then plateaued for 7 days. During the 7 days the cells continued to express E-cadherin; however, tangential AJs typical for cells with stable cell-cell adhesion, changed into radial AJs. The radial AJs continued to accumulate E-cadherin during 24‑72 hours after tetracycline removal. As a result of SNAI1 activation, the cells underwent epithelial-mesenchymal transition (EMT) and became migratory. On a two-dimensional substrate, cells exhibited both individual and collective migration. As the tetracycline washout period progressed, the fraction of the cells capable of migrating through migration chamber membranes increased; on the contrary, cells’ ability to invade an epithelial monolayer decreased. These results demonstrate that retaining a hybrid epithelial/mesenchymal phenotype and accumulation of E-cadherin in AJs during early stages of EMT do not impede disruption of stable cell-cell adhesion and cells’ acquisition of migratory activity.
About the Authors
I. Y. Zhitnyak
FSBI «N.N. Blokhin National Medical Research Center of Oncology» of the Ministry of Health of the Russian Federation
Russian Federation
Russian Federation
24, Kashirskoye Shosse, 115478-Moscow, Russia
PhD, Research scientist in the Laboratory of the Mechanisms of Carcinogenesis, FSBI «N.N. Blokhin National Medical Research Center of Oncology» of the Ministry of Health of the Russian Federation
Author ID (Scopus): 35849650500.
N. I. Litovka
FSBI «N.N. Blokhin National Medical Research Center of Oncology» of the Ministry of Health of the Russian Federation
Russian Federation
Russian Federation
24, Kashirskoye Shosse, 115478-Moscow, Russia
Technician in the Laboratory of the Mechanisms of Carcinogenesis, FSBI «N.N. Blokhin National Medical Research Center of Oncology» of the Ministry of Health of the Russian Federation
S. N. Rubtsova
FSBI «N.N. Blokhin National Medical Research Center of Oncology» of the Ministry of Health of the Russian Federation
Russian Federation
Russian Federation
24, Kashirskoye Shosse, 115478-Moscow, Russia
PhD, Research scientist in the Laboratory of the Mechanisms of Carcinogenesis, FSBI «N.N. Blokhin National Medical Research Center of Oncology» of the Ministry of Health of the Russian Federation
Author ID (Scopus): 15754557800
N. A. Gloushankova
FSBI «N.N. Blokhin National Medical Research Center of Oncology» of the Ministry of Health of the Russian Federation
Russian Federation
Russian Federation
24, Kashirskoye Shosse, 115478-Moscow, Russia
DSc, Head of the Laboratory of the Mechanisms of Carcinogenesis, FSBI «N.N. Blokhin National Medical Research Center of Oncology» of the Ministry of Health of the Russian Federation
Author ID (Scopus): 6507200005
References
1. Nieto M.A. The snail superfamily of zinc-finger transcription factors. Nat Rev Mol Cell Biol. 2002 Mar; 3(3): 155–66. doi: 10.1038/nrm757.
2. Cano A., Pérez-Moreno M.A., Rodrigo I., Locascio A., Blanco M.J., del Barrio M.G., Portillo F., Nieto M.A. The transcription factor Snail controls epithelial mesenchymal transitions by repressing E-cadherin expression. Nat Cell Biol. 2000 Feb; 2(2): 76–83. doi: 10.1038/35000025.
3. Batlle E., Sancho E., Francí C., Domínguez D., Monfar M., Baulida J., García De Herreros A. The transcription factor snail is a repressor of E-cadherin gene expression in epithelial tumour cells. Nat Cell Biol. 2000 Feb; 2(2): 84–9. doi: 10.1038/35000034.
4. Ikenouchi J., Matsuda M., Furuse M., Tsukita S. Regulation of tight junctions during the epithelium-mesenchyme transition: direct repression of the gene expression of claudins/occludin by Snail. J Cell Sci. 2003 May 15; 116: 1959–67. doi: 10.1242/jcs.00389.
5. Beach S., Tang H., Park S., Dhillon A.S., Keller E.T., Kolch W., Yeung K.C. Snail is a repressor of RKIP transcription in metastatic prostate cancer cells. Oncogene. 2008 Apr 3; 27(15): 2243–8. doi: 10.1038/sj.onc.1210860.
6. Barrallo-Gimeno A., Nieto M.A. The Snail genes as inducers of cell movement and survival: implications in development and cancer. Development. 2005 Jul; 132(14): 3151–61. doi: 10.1242/dev.01907.
7. Kaufhold S., Bonavida J. Central role of Snail1 in the regulation of EMT and resistance in cancer: a target for therapeutic intervention. J Exp Clin Cancer Res. 2014 Aug 2; 33: 62. doi: 10.1186/s13046-014-0062-0.
8. Nieto M.A., Huang R.Y.J., Jackson R.A., Thiery J.P. EMT: 2016. Cell. 2016 Jun 30; 166(1): 21–45. doi: 10.1016/j.cell.2016.06.028.
9. Herrera A., Herrera M., Alba-Castellón L., Silva J., García V., Loubat-Casanovas J., Alvarez-Cienfuegos A., García J.M., Rodriguez R., Gil B., Citores M.J., Larriba M.J., Casal J.I., Garcia de Herreros A., Bonilla F., Peña C. Protumorigenic effects of Snail- expression fibroblasts on colon cancer cells. Int J Cancer. 2014 Jun 15; 134(12): 2984- 904–9. doi: 10.1002/ijc.28613.
10. Yatskou M., Novikov E., Vetter G., Muller A., Barillot E., Vallar L., Friederich E. Advanced spot quality analysis in two-colour microarray experiments. BMC Res Notes. 2008 Sep 17; 1: 80. doi: 10.1186/1756-0500-1-80.
11. Rubtsova S.N., Zhitnyak I.Y., Gloushankova N.A. A Novel Role of E-Cadherin- Based Adherens Junctions in Neoplastic Cell Dissemination. PLoS One. 2015 Jul 24; 10(7): e0133578. doi: 10.1371/journal.pone.0133578.
12. Vetter G., Le Bechec A., Muller J., Moes M., Yatskou M., Al Tanoury Z., Poch O., Vallar L., Friederich E. Time-resolved analysis of transcriptional events during SNAI1- triggered epithelial to mesenchymal transition. Biochem Biophys Res Comm. 2009. Aug 7; 385(4): 485–91. doi: 10.1016/j.bbrc.2009.05.025.
13. Ayollo D.V., Zhitnyak I.Y., Vasiliev J.M., Gloushankova N.A. Rearrangements of the Actin Cytoskeleton and E-Cadherin–Based Adherens Junctions Caused by Neoplasic Transformation Change Cell–Cell Interactions. PLoS One. 2009 Nov 30; 4(11): e8027. doi: 10.1371/journal.pone.0008027.
14. Zhitnyak I.Yu., Gloushankova N.A. Morphology, Cell-Cell Interactions, and Migratory Activity of IAR-2 Epithelial Cells Transformed with the RAS Oncogene: Contribution of Cell Adhesion Protein E-Cadherin. Ontogenez. 2011 Nov-Dec; 42(6): 453–64.
15. Odero-Marah V.A., Khalkhali-Ellis Z., Chunthapong J., Amir S., Seftor R.E., Seftor E.A., Hendrix M.J. Maspin regulates different signaling pathways for motility and adhesion in aggressive breast cancer cells. Cancer Biol Ther. 2003 Jul-Aug; 2(4): 398–403.
16. Neal C.L., Henderson V., Smith B.N., McKeithen D., Graham T., Vo B.T., Odero- Marah V.A. Snail transcription factor negatively regulates maspin tumor suppressor in human prostate cancer cells. BMC Cancer. 2012 Aug 2; 12: 336. doi: 10.1186/1471-2407-12-336.
Review
For citations:
Zhitnyak I.Y., Litovka N.I., Rubtsova S.N., Gloushankova N.A. INDUCTION OF EPITHELIAL-TO-MESENCHYMAL TRANSITION IN MCF-7-SNAI1 CELLS LEADS TO REORGANIZATION OF ADHERENS JUNCTIONS AND ACQUISITION OF MIGRATORY ACTIVITY. Siberian journal of oncology. 2018;17(4):24-29. https://doi.org/10.21294/1814-4861-2018-17-4-24-29
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