Inhibitory effect of vector-mediated RNAi technique on expression and function of vitamin D3 receptor in human osteosarcoma cell line HOS-8603
- Author:
Ye ZHANG
1
Author Information
1. Department of Pathophysiology
- Publication Type:Journal Article
- From:
Academic Journal of Second Military Medical University
2006;27(10):1076-1080
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To construct an expression vector containing RNA interfering (RNAi) sequence targeting human vitamin D3 receptor (VDR) protein gene, and to investigate its inhibitory effect on the expression and function of VDR in a human osteosarcoma cell line. Methods: Two small interfering RNA sequences targeting VDR were designed by online screening and were cloned into the expression vector (pSilencer-2. 1-U6). Then the products, pSilencer-2. 1-U6-VDR1 and pSilencer-2. 1-U6-VDR2, were separately transfected into a human osteosarcoma cell line (HOS-8603) by liposome. Western blot was employed to determine their effect on VDR protein expression and 1,25(OH)2D3-induced phosphorylation of PKC. Cell counting and MTT were used to evaluate their influence on the inhibitory effects of 1,25(OH)2D3 on HOS-8603 cells proliferation. Results: Compared to HOS-8603 cells transfected with control vectors, those transfected with pSilencer2. 1-U6-VDR1 or pSilencer2. 1-U6-VDR2 had significant decreases in the expression of VDR protein, in the inhibitory effects of 1,25(OH)2D3 on the cell proliferation, and in quick phosphorylation of PKC induced by 1,25(OH)2D3. Conclusion: The vectors constructed in the present study (pSilencer2. 1-U6-VDR1 and pSilencer2. 1-U6-VDR2) can significantly suppress the VDR expression in HOS-8603 cells, and block some biological activities of 1,25(OH)2D3 mediated by VDR, which paves a way for further study on the biological functions of VDR and vitamin D3.
