title: The adaptor protein p62 is involved in RANKL-induced autophagy and osteoclastogenesis.
abstract: Previous studies have implicated autophagy in osteoclast differentiation. The aim of this study was to investigate the potential role of p62, a characterized adaptor protein for autophagy, in RANKL-induced osteoclastogenesis. Real-time quantitative PCR and western blot analyses were used to evaluate the expression levels of autophagy-related markers during RANKL-induced osteoclastogenesis in mouse macrophage-like RAW264.7 cells. Meanwhile, the potential relationship between p62/LC3 localization and F-actin ring formation was tested using double-labeling immunofluorescence. Then, the expression of p62 in RAW264.7 cells was knocked down using small-interfering RNA (siRNA), followed by detecting its influence on RANKL-induced autophagy activation, osteoclast differentiation, and F-actin ring formation. The data showed that several key autophagy-related markers including p62 were significantly altered during RANKL-induced osteoclast differentiation. In addition, the expression and localization of p62 showed negative correlation with LC3 accumulation and F-actin ring formation, as demonstrated by western blot and immunofluorescence analyses, respectively. Importantly, the knockdown of p62 obviously attenuated RANKL-induced expression of autophagy- and osteoclastogenesis-related genes, formation of TRAP-positive multinuclear cells, accumulation of LC3, as well as formation of F-actin ring. Our study indicates that p62 may play essential roles in RANKL-induced autophagy and osteoclastogenesis, which may help to develop a novel therapeutic strategy against osteoclastogenesis-related diseases.
authors.affiliation: State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) & Key Laboratory of Oral Biomedicine Ministry of Education, School & Hospital of Stomatology (RFL, GC, JGR, WZ, ZXW, BL, YZ, YFZ) Wuhan University, Wuhan, ChinaDepartment of Oral and Maxillofacial Surgery, School & Hospital of Stomatology (GC, ZXW, BL, YFZ) Wuhan University, Wuhan, ChinaDepartment of Prosthodontics, School & Hospital of Stomatology (YZ) Wuhan University, Wuhan, China.
authors.forename: Rui-Fang
authors.initials: RF
authors.lastname: Li
authors.affiliation: State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) & Key Laboratory of Oral Biomedicine Ministry of Education, School & Hospital of Stomatology (RFL, GC, JGR, WZ, ZXW, BL, YZ, YFZ) Wuhan University, Wuhan, ChinaDepartment of Oral and Maxillofacial Surgery, School & Hospital of Stomatology (GC, ZXW, BL, YFZ) Wuhan University, Wuhan, ChinaDepartment of Prosthodontics, School & Hospital of Stomatology (YZ) Wuhan University, Wuhan, China.
authors.forename: Gang
authors.initials: G
authors.lastname: Chen
authors.affiliation: State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) & Key Laboratory of Oral Biomedicine Ministry of Education, School & Hospital of Stomatology (RFL, GC, JGR, WZ, ZXW, BL, YZ, YFZ) Wuhan University, Wuhan, ChinaDepartment of Oral and Maxillofacial Surgery, School & Hospital of Stomatology (GC, ZXW, BL, YFZ) Wuhan University, Wuhan, ChinaDepartment of Prosthodontics, School & Hospital of Stomatology (YZ) Wuhan University, Wuhan, China.
authors.forename: Jian-Gang
authors.initials: JG
authors.lastname: Ren
authors.affiliation: State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) & Key Laboratory of Oral Biomedicine Ministry of Education, School & Hospital of Stomatology (RFL, GC, JGR, WZ, ZXW, BL, YZ, YFZ) Wuhan University, Wuhan, ChinaDepartment of Oral and Maxillofacial Surgery, School & Hospital of Stomatology (GC, ZXW, BL, YFZ) Wuhan University, Wuhan, ChinaDepartment of Prosthodontics, School & Hospital of Stomatology (YZ) Wuhan University, Wuhan, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Zhang
authors.affiliation: State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) & Key Laboratory of Oral Biomedicine Ministry of Education, School & Hospital of Stomatology (RFL, GC, JGR, WZ, ZXW, BL, YZ, YFZ) Wuhan University, Wuhan, ChinaDepartment of Oral and Maxillofacial Surgery, School & Hospital of Stomatology (GC, ZXW, BL, YFZ) Wuhan University, Wuhan, ChinaDepartment of Prosthodontics, School & Hospital of Stomatology (YZ) Wuhan University, Wuhan, China.
authors.forename: Zhong-Xing
authors.initials: ZX
authors.lastname: Wu
authors.affiliation: State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) & Key Laboratory of Oral Biomedicine Ministry of Education, School & Hospital of Stomatology (RFL, GC, JGR, WZ, ZXW, BL, YZ, YFZ) Wuhan University, Wuhan, ChinaDepartment of Oral and Maxillofacial Surgery, School & Hospital of Stomatology (GC, ZXW, BL, YFZ) Wuhan University, Wuhan, ChinaDepartment of Prosthodontics, School & Hospital of Stomatology (YZ) Wuhan University, Wuhan, China.
authors.forename: Bing
authors.initials: B
authors.lastname: Liu
authors.affiliation: State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) & Key Laboratory of Oral Biomedicine Ministry of Education, School & Hospital of Stomatology (RFL, GC, JGR, WZ, ZXW, BL, YZ, YFZ) Wuhan University, Wuhan, ChinaDepartment of Oral and Maxillofacial Surgery, School & Hospital of Stomatology (GC, ZXW, BL, YFZ) Wuhan University, Wuhan, ChinaDepartment of Prosthodontics, School & Hospital of Stomatology (YZ) Wuhan University, Wuhan, China zhao_yi@whu.edu.cn.
authors.forename: Yi
authors.initials: Y
authors.lastname: Zhao
authors.affiliation: State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) & Key Laboratory of Oral Biomedicine Ministry of Education, School & Hospital of Stomatology (RFL, GC, JGR, WZ, ZXW, BL, YZ, YFZ) Wuhan University, Wuhan, ChinaDepartment of Oral and Maxillofacial Surgery, School & Hospital of Stomatology (GC, ZXW, BL, YFZ) Wuhan University, Wuhan, ChinaDepartment of Prosthodontics, School & Hospital of Stomatology (YZ) Wuhan University, Wuhan, China.
authors.forename: Yi-Fang
authors.initials: YF
authors.lastname: Zhao
mesh_terms: D000199:Actins / Q000032:analysis / Q000378:metabolism; D000818:Animals; D001343:Autophagy; D002454:Cell Differentiation; D002460:Cell Line; D055785:Gene Knockdown Techniques; D051379:Mice; D010010:Osteoclasts / Q000166:cytology* / Q000378:metabolism; D053245:RANK Ligand / Q000032:analysis / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D051758:Transcription Factor TFIIH; D014157:Transcription Factors / Q000032:analysis / Q000235:genetics / Q000378:metabolism*
chemical_list: D000199:Actins; C490981:Gtf2h1 protein, mouse; D053245:RANK Ligand; D014157:Transcription Factors; D051758:Transcription Factor TFIIH
keywords: RANKL; RAW264.7 cells; autophagy; osteoclastogenesis; p62