High-density seeding affects transforming growth factor-beta1-induced epithelial-mesenchymal transition of HaCaT cells

doi:10.3969/j.issn.2095-4344.2013.28.015

Abstract

Abstract:

BACKGROUND: The cell density is one of the factors involved in the state of cell differentiation, and the effect of cell density on transforming growth factor-β1-induced epithelial-mesenchymal transition of HaCaT cells is still unclear.
OBJECTIVE: To observe the effect of cell density on transforming growth factor-β1-induced epithelial-mesenchymal transition of HaCaT cells.
METHODS: HaCaT cells was seeded in 6-well plates at low density of 103/cm2 and high density of 105/cm2, and then treated by 2 μg/L transforming growth factor-β1 for 48 hours, thereafter observed the changes in cell morphology. The transcription levels of epithelial cadherin, tight junction protein-1, vimentin, neuronal-cadherin were detected by real-time PCR, and expression levels of epithelial cadherin and vimentin were detected by Western blot.
RESULTS AND CONCLUSION: Cell gap of HaCaT cells grew larger after treated with transforming growth factor-β1 for 48 hours, and cell morphology was long spindle rather than polygonal in low-density group, while in high-density group without obvious morphological changes. The real-time PCR showed that the transcriptions of epithelial cell marker epithelial cadherin and tight junction protein-1 were suppressed when compared with the control group (P < 0.05), and the decreasing deree in the high-density group was higher than that in the low-density group (P < 0.05), mesenchymal cell marker neuronal-cadherin and vimentin were upregulated in the high-density group and the low-density group when compared with the control group (P < 0.05), and there were no significant differences between high-density group and low-density group. Western blot results verified the changes of neuronal-cadherin and vimentin expression level. These results suggest that the high seeding density can inhibit transforming growth factor-β1-induced epithelial-mesenchymal transition of HaCaT cells.

Key words: tissue construction, tissue construction and bioactive factors, transforming growth factor-β1, HaCaT cells, bioactive factors, cell density, epithelial-mesenchymal transition, National 973 Project of China

CLC Number:

R318

Zhang Dian-bao, Wang Zhe, Shi Ping, Pang Xi-ning. High-density seeding affects transforming growth factor-beta1-induced epithelial-mesenchymal transition of HaCaT cells[J]. Chinese Journal of Tissue Engineering Research, 2013, 17(28): 5191-5197.

Figures/Tables 5

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