title: microRNA-143 is associated with the survival of ALDH1+CD133+ osteosarcoma cells and the chemoresistance of osteosarcoma.
abstract: This study investigated the role of miR-143 in the chemoresistance of osteosarcoma tumor cells and the associated mechanisms. Real-time PCR was used to measure miR-143 levels. Western blot was used to detect protein expression. Cell proliferation was analyzed by MTT assay and Matrigel colony formation assay. Forced miR-143 expression was established by adenoviral vector infection. Cell death was detected by Hoechst33342 staining. Loss of miR-143 expression was observed in osteosarcomas, which correlated with shorter survival of patients with osteosarcomas underlying chemotherapy. In chemoresistant SAOS-2 and U2OS osteosarcomas cells, miR-143 levels were significantly downregulated and accompanied by increases in ATG2B, Bcl-2, and/or LC3-II protein levels, high rate of ALDH1(+)CD133(+) cells, and an increase in Matrigel colony formation ability. H2O2 upregulated p53 and miR-143, but downregulated ATG2B, Bcl-2, and LC3-I expression in U2OS cells (wild-type p53) but not in SAOS-2 (p53-null) cells. Forced miR-143 expression significantly reversed chemoresistance as well as downregulation of ATG2B, LC3-I, and Bcl-2 expression in SAOS-2- and U2OS-resistant cells. Forced miR-143 expression significantly inhibited tumor growth in xenograft SAOS-2-Dox and U2OS-Dox animal models. Loss of miR-143 expression is associated with poor prognosis of patients with osteosarcoma underlying chemotherapy. The chemoresistance of osteosarcoma tumor cells to doxorubicin is associated with the downregulation of miR-143 expression, activation of ALDH1(+)CD133(+) cells, activation of autophagy, and inhibition of cell death. miR-143 may play a crucial role in the chemoresistance of osterosarcoma tumors.
authors.affiliation: Department of Orthopedics, Third Xiangya Hospital, Central South University, Changsha 410013, China.
authors.forename: Jiahui
authors.initials: J
authors.lastname: Zhou
authors.affiliation: Department of Orthopedics, Third Xiangya Hospital, Central South University, Changsha 410013, China wusong08@hotmail.com.
authors.forename: Song
authors.initials: S
authors.lastname: Wu
authors.affiliation: Hepatobiliary and Enteric Surgery Research Center, Xiangya Hospital, Central South University, Changsha 410008, China.
authors.forename: Yuxiang
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Hepatobiliary and Enteric Surgery Research Center, Xiangya Hospital, Central South University, Changsha 410008, China.
authors.forename: Jingfeng
authors.initials: J
authors.lastname: Zhao
authors.affiliation: Department of Orthopedics, Third Xiangya Hospital, Central South University, Changsha 410013, China.
authors.forename: Kexiang
authors.initials: K
authors.lastname: Zhang
authors.affiliation: Department of Orthopedics, Third Xiangya Hospital, Central South University, Changsha 410013, China.
authors.forename: Jianlong
authors.initials: J
authors.lastname: Wang
authors.affiliation: Department of Orthopedics, Third Xiangya Hospital, Central South University, Changsha 410013, China.
authors.forename: Shijie
authors.initials: S
authors.lastname: Chen
mesh_terms: D000071916:AC133 Antigen; D000080924:Aldehyde Dehydrogenase 1 Family; D000818:Animals; D015703:Antigens, CD / Q000096:biosynthesis; D015153:Blotting, Western; D001859:Bone Neoplasms / Q000235:genetics* / Q000401:mortality / Q000473:pathology; D045744:Cell Line, Tumor; D019008:Drug Resistance, Neoplasm / Q000235:genetics*; D015972:Gene Expression Regulation, Neoplastic; D006023:Glycoproteins / Q000096:biosynthesis; D064593:Heterografts; D006801:Humans; D007527:Isoenzymes / Q000096:biosynthesis; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D035683:MicroRNAs / Q000096:biosynthesis*; D012516:Osteosarcoma / Q000235:genetics* / Q000401:mortality / Q000473:pathology; D010455:Peptides; D060888:Real-Time Polymerase Chain Reaction; D050697:Retinal Dehydrogenase / Q000096:biosynthesis
chemical_list: D000071916:AC133 Antigen; D015703:Antigens, CD; D006023:Glycoproteins; D007527:Isoenzymes; C542359:MIRN143 microRNA, human; D035683:MicroRNAs; C000606023:PROM1 protein, human; D010455:Peptides; C000606024:Prom1 protein, mouse; D000080924:Aldehyde Dehydrogenase 1 Family; C510223:ALDH1A1 protein, human; C515028:ALDH1A1 protein, mouse; D050697:Retinal Dehydrogenase
keywords: ALDH1+CD133+ cell; apoptosis; autophagy; miR-143; osteosarcoma