Related Experiment Video
Updated: Aug 1, 2026
Isolation of Mesenchymal Stem Cells from Human Alveolar Periosteum and Effects of Vitamin D on Osteogenic Activity of Periosteum-derived Cells
Published on: May 4, 2018
Effects of 1,25(OH)2D3, EB1089, and analog V on PTHrP production, PTHrP mRNA expression and cell growth in SCC 2/88
S Kunakornsawat1, T J Rosol, C C Capen
1Department of Veterinary Biosciences, College of Veterinary Medicine, The Ohio State University, Columbus 43210-1092, USA.
Background:
We investigated the effects of 1,25(OH)2D3 and selected analogs on canine squamous carcinoma cells (SCC 2/88) and tested whether these compounds could effectively decrease proliferation, induce differentiation, and inhibit PTHrP production and PTHrP mRNA expression.
Materials And Methods:
SCC 2/88 cells were cultured and treated with three substrates. The media were collected for PTHrP immunoradiometric assay. The cells were analyzed for DNA concentration and PTHrP mRNA expression by Northern blot analysis, involucrin by Western blot analysis and 1,25(OH)2D3-receptor (VDR) and PTHrP by immunohistochemistry.
Results:
The SCC 2/88 cells were stained positively for VDR and PTHrP by immunohistochemistry. 1,25(OH)2D3 and its analogs inhibited cell growth and stimulated differentiation in a dose-dependent manner. All three substrate-treated groups had significantly increased PTHrP secretion at 10(-7) M. Cells treated with 1,25(OH)2D3 at 10(-7) M had 2- to 4-fold increased PTHrP mRNA expression at 12 and 24 hours compared to the vehicle-treated controL PTHrP mRNA in cells treated with TGF-beta (1.5 ng/ml) was increased 7- to 17-fold at 6, 12 and 24 hours compared to the vehicle-treated controL PTHrP mRNA expression was reduced by 0.5- to 2-fold in cells treated with 1,25(OH)2D3 at 10(-7) M and TGF-beta (1.5 ng/ml) together compared to cells treated with TGF-beta alone.
Conclusion:
1,25(OH)2D3, EB1089, and analog V inhibited SCC 2/88 growth and induced differentiation in a dose-dependent manner, but did not inhibit PTHrP production. 1,25(OH)2D3 treatment led to increased PTHrP mRNA expression and reduced the stimulatory effect of TGF-beta on PTHrP mRNA expression in SCC 2/88 cells.
Insights
Vitamin D analogs like 1,25(OH)2D3 inhibit canine squamous cell carcinoma growth and induce differentiation. However, they increase parathyroid hormone-related peptide (PTHrP) mRNA expression, not inhibit its production.
Area of Science:
- Oncology
- Endocrinology
- Molecular Biology
Background:
- Investigated the effects of 1,25(OH)2D3 and analogs on canine squamous carcinoma cells (SCC 2/88).
- Assessed the potential of these compounds to decrease proliferation, induce differentiation, and inhibit PTHrP production and mRNA expression.
Purpose of the Study:
- To determine the impact of 1,25(OH)2D3 and its analogs on canine SCC cell behavior.
- To evaluate the modulation of PTHrP production and gene expression by these vitamin D compounds.
Main Methods:
- SCC 2/88 cells were cultured and treated with 1,25(OH)2D3 and analogs.
- PTHrP levels, DNA concentration, and mRNA expression were analyzed using immunoradiometric assay, Northern blot, and Western blot.
- Cellular expression of VDR and PTHrP was examined via immunohistochemistry.
Main Results:
- 1,25(OH)2D3 and analogs inhibited SCC 2/88 cell growth and stimulated differentiation dose-dependently.
- PTHrP secretion increased significantly at 10(-7) M for all substrate-treated groups.
- 1,25(OH)2D3 treatment increased PTHrP mRNA expression and partially reduced TGF-beta's stimulatory effect on it.
Conclusions:
- 1,25(OH)2D3, EB1089, and analog V inhibit SCC 2/88 growth and induce differentiation without inhibiting PTHrP production.
- 1,25(OH)2D3 increases PTHrP mRNA expression in SCC 2/88 cells.
- Vitamin D compounds modulate TGF-beta's effect on PTHrP mRNA expression.
