Inhibitory effect of vector-mediated RNAi technique on expression and function of vitamin D_3 receptor in human osteosarcoma cell line HOS-8603
- VernacularTitle:利用载体介导的RNA干扰技术抑制人骨肉瘤细胞HOS-8603内维生素D_3受体的表达及功能
- Author:
Ye ZHANG
- Publication Type:Journal Article
- Keywords:
RNA interference;
osteosarcoma;
receptors,vitamin D_3
- From:
Academic Journal of Second Military Medical University
2000;0(10):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To construct an expression vector containing RNA interfering(RNAi) sequence targeting human vitamin D_3 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)_2D_3-induced phosphorylation of PKC.Cell counting and MTT were used to evaluate their influence on the inhibitory effects of 1,25(OH)_2D_3 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)_2D_3 on the cell proliferation,and in quick phosphorylation of PKC induced by 1,25(OH)_2D_3.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)_2D_3 mediated by VDR, which paves a way for further study on the biological functions of VDR and vitamin D_3.
