Effect of tumor necrosis factor receptor associated factor 6 gene silencing by small interfering RNA on proliferation of odontoblast cells
10.3760/cma.j.issn.1002-0098.2014.07.010
- VernacularTitle:肿瘤坏死因子受体相关因子6基因沉默对成牙本质细胞增殖能力的影响
- Author:
Xia LI
1
;
Mingzhen XIAO
;
Qing YU
;
Bin ZHAO
;
Weixing LI
Author Information
1. 山西医科大学口腔医院牙体牙髓病科
- Keywords:
RNA interfering;
Transfection;
Cell proliferation;
Tumor necrosis factor receptor associated factor 6
- From:
Chinese Journal of Stomatology
2014;49(7):428-433
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effect of small interfering RNA(siRNA)-mediated inhibition of tumor necrosis factor receptor associated factor 6(TRAF6) gene on murine odontoblast-like cell line,MDPC-23 cell and the effect of TRAF6 on MDPC-23 cell proliferation.Methods The vectors expressing siRNA against TRAF6 were constructed and introduced into MDPC-23 cell with lipofectin,and the cell line with stable expression of siRNA of TRAF6 was obtained by G418 screening and colony culture.Reverse transcription-PCR(RT-PCR) and Western blotting were performed to detect the expression of TRAF6.The proliferation of transfected MDPC-23 cell was investigated through methabenzthiazuron(MTF)and flow cytometry(FCM) assay.Results The positive single colony was screened out,and was found to express siRNA against TRAF6 effectively because both TRAF6 mRNA and protein relative expression were significantly decreased in the experimental group(pSUPER-TRAF6siRNA:mRNA 0.163 ± 0.008,protein 0.215±0.006) compared with controls(pSUPER:mRNA 0.778±0.017,protein 0.964±0.007 (P ＜ 0.001).The A value of treated pSUPER-TRAF6siRNA cells (3 d:0.46±0.03,5 d:1.35±0.06) was increased compared with controls(P＜0.01).The result of proliferation index(PrI) was also increased compared with controls [pSUPER-TRAF6siRNA:(24.1 ± 2.2)％ ; pSUPER(11.2 ± 1.0)％ ; control(10.5 ± 0.7)％,P＜0.01].Conclusions The transcription and expression of TRAF6 gene were inhibited.The proliferation ability was increased in MDPC-23 cells by the constructed pSUPER-TRAF6siRNA vector.It may further influence the formation and repair of dentin,and may be involved in the regulation of normal tooth eruption and process of dentin repair after injury.
