title: miR-223 contributes to the AGE-promoted apoptosis via down-regulating insulin-like growth factor 1 receptor in osteoblasts.
abstract: Advanced glycation end products (AGEs) have been confirmed to induce bone quality deterioration in diabetes mellitus (DM), and to associate with abnormal expression of miRNAs in DM patients or in vitro Recently, miRNAs have been recognized to mediate the onset or progression of DM. In the present study, we investigated the regulation on miR-223 level by AGE-BSA treatment in osteoblast-like MC3T3-E1 cells, with real-time quantitative PCR assay. And then we examined the inhibition of insulin-like growth factor 1 receptor (IGF-1R) expression by miR-223, via targeting of the 3' UTR of IGF-1R with real-time quantitative PCR, western blotting and luciferase reporter assay. Then we explored the regulation of miR-223 and IGF-1R levels, via the lentivirus-mediated miR-223 inhibition and IGF-1R overexpression in the AGE-BSA-induced apoptosis in MC3T3-E1 cells. It was demonstrated that AGE-BSA treatment with more than 100 μg/ml significantly up-regulated miR-223 level, whereas down-regulated IGF-1R level in MC3T3-E1 cells. And the up-regulated miR-223 down-regulated IGF-1R expression in both mRNA and protein levels, via targeting the 3' UTR of IGF-1R Moreover, though the AGE-BSA treatment promoted apoptosis in MC3T3-E1 cells, the IGF-1R overexpression or the miR-223 inhibition significantly attenuated the AGE-BSA-promoted apoptosis in MC3T3-E1 cells. In summary, our study recognized the promotion of miR-223 level by AGE-BSA treatment in osteoblast-like MC3T3-E1 cells. The promoted miR-223 targeted IGF-1R and mediated the AGE-BSA-induced apoptosis in MC3T3-E1 cells. It implies that miR-223 might be an effective therapeutic target to antagonize the AGE-induced damage to osteoblasts in DM.
authors.affiliation: Department of Orthopedics, Sun Yat-Sen Memorial Hospital, Sun Yat-sen University, Guangzhou 510120, China Department of Orthopedics, Zhuhai People's Hospital, Zhuhai 519000, China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Qin
authors.affiliation: Department of Orthopedics, Sun Yat-Sen Memorial Hospital, Sun Yat-sen University, Guangzhou 510120, China.
authors.forename: Jichao
authors.initials: J
authors.lastname: Ye
authors.affiliation: Department of Orthopedics, Sun Yat-Sen Memorial Hospital, Sun Yat-sen University, Guangzhou 510120, China.
authors.forename: Peng
authors.initials: P
authors.lastname: Wang
authors.affiliation: Department of Orthopedics, Sun Yat-Sen Memorial Hospital, Sun Yat-sen University, Guangzhou 510120, China.
authors.forename: Liangbin
authors.initials: L
authors.lastname: Gao
authors.affiliation: Department of Orthopedics, Zhuhai People's Hospital, Zhuhai 519000, China.
authors.forename: Suwei
authors.initials: S
authors.lastname: Wang
authors.affiliation: Department of Orthopedics, Sun Yat-Sen Memorial Hospital, Sun Yat-sen University, Guangzhou 510120, China huiyoshen@163.com.
authors.forename: Huiyong
authors.initials: H
authors.lastname: Shen
mesh_terms: D020413:3' Untranslated Regions; D000818:Animals; D017209:Apoptosis / Q000187:drug effects* / Q000235:genetics; D002460:Cell Line; D015536:Down-Regulation / Q000187:drug effects*; D017127:Glycation End Products, Advanced / Q000378:metabolism / Q000493:pharmacokinetics*; D051379:Mice; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D010006:Osteoblasts / Q000166:cytology / Q000378:metabolism*; D017526:Receptor, IGF Type 1 / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D020413:3' Untranslated Regions; D017127:Glycation End Products, Advanced; C544877:MIRN223 microRNA, mouse; D035683:MicroRNAs; D017526:Receptor, IGF Type 1
keywords: advanced glycation end products (AGEs); apoptosis; insulin-like growth factor 1 receptor (IGF-1R); miR-223; osteoblasts