High-density seeding affects transforming growth factor-beta1-induced epithelial-mesenchymal transition of HaCaT cells
10.3969/j.issn.2095-4344.2013.28.015
- VernacularTitle:高密度接种对转化生长因子β1诱导上皮间充质转化的影响**
- Author:
Dianbao ZHANG
;
Zhe WANG
;
Ping SHI
;
Xining PANG
- Publication Type:Journal Article
- Keywords:
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
- From:
Chinese Journal of Tissue Engineering Research
2013;(28):5191-5197
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUND: The cel density is one of the factors involved in the state of cel differentiation, and the effect of cel density on transforming growth factor-β1-induced epithelial-mesenchymal transition of HaCaT cells is stil unclear. OBJECTIVE: To observe the effect of cel density on transforming growth factor-β1-induced epithelial-mesenchymal transition of HaCaT cells. METHODS: HaCaT cells was seeded in 6-wel plates at low density of 103/cm2 and high density of 105/cm2 then treated by 2 μg/L transforming growth factor-β1 for 48 hours, thereafter observed the changes in cel 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: Cel gap of HaCaT cells grew larger after treated with transforming growth factor-β1 for 48 hours, and cel 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 cel 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 cel 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.
