Effect of silencing human RRM 2 by specific siRNA on the biological behavior of human osteosarcoma cell
10.3760/cma.j.issn.1008-6706.2013.17.003
- VernacularTitle:特异沉默人核糖核苷酸还原酶M2基因对人骨肉瘤细胞生物学特性的影响
- Author:
Teng LIAO
;
Zhong WANG
;
Yiping WANG
;
Wentao HUANG
;
Hao SHEN
- Publication Type:Journal Article
- Keywords:
RNA interference;
Gene,RRM2;
Osteosarcoma
- From:
Chinese Journal of Primary Medicine and Pharmacy
2013;20(17):2566-2569
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effect of small interference RNA(siRNA) targeting RRM2 on the biological behavior of human osteosarcoma cell line Saos-2 and the molecular mechanisms.Methods RRM2 expression was knocked down in human osteosarcama cell line Saos-2 by RRM2 siRNA.The expression of RRM2 mRNA and protein was determined in human osteosarcoma cell line Saos-2 and human osteoblast-like cell line hFOB1.19 by real time-PCR and Western blot.The cell proliferation was detected by CCK-8.The migration was observed by using transwell system.The apoptotic rate was observed by ELISA.The expression of CyclinD1 and Bcl-2 proteins were detected by Western blot.Results The expression of RRM2 mRNA and protein was higher in Saos-2 than in hFOB1.19.siRRM2 could down-regulate the expression of RRM2 in Saos-2 cells in a time-and concentration-dependent manner.CCK-8 assay showed that si-RRM2 could inhibit the proliferation ability of Saos-2 cells in a time-and concentrationdependent manner,but had no effect on the proliferation of hFOB1.19 cells.Transwell assay indicated that si-RRM2 could inhibit the migration of Saos-2 cells.si-RRM2 combined with adriamycin could increase the apoptosis of Saos-2 cells.Western blot showed that the expression of Cyclin D1 and Bcl-2 were decreased by silencing RRM2.Condusion RRM2 overexpression maybe associate with the osteosarcoma cells proliferation and migration and suppression of its function is a potential therapeutic strategy in osteosarcoma.
