title: MicroRNA-184 Modulates Doxorubicin Resistance in Osteosarcoma Cells by Targeting BCL2L1.
abstract: BACKGROUND Early metastasis of osteosarcoma (OS) is highly lethal and responds poorly to drug and radiation therapies. MicroRNAs (miRNAs) are a class of small noncoding RNAs that modulate gene expression at the post-transcriptional level. However, the detailed functions of specific miRNAs are not entirely understood. The aim of the present study was to investigate the role of miR-184 as a mediator of drug resistance in human osteosarcoma. MATERIAL AND METHODS qRT-PCR was used to analyze the expression level of miR-184 in OS cell line U-2 OS and MG-63 treated with doxorubicin. MiR-184 agomir or miR-184 antagomir was transferred into cells to regulated miR-184. The target of miR-184 was predicted by TargetScan and confirmed by luciferase reporter assay. Bcl-2-like protein 1 (BCL2L1) expression was detected by Western blot. Cell apoptosis was determined by Annexin V staining and analysis by flow cytometry. RESULTS Doxorubicin induced time-dependent expression of miR-184 in OS cell line U-2 OS and MG-63. Luciferase reporter assay identiﬁed BCL2L1 as the direct target gene of miR-184. Furthermore, doxorubicin reduced BCL2L1 expression, which was reversed by miR-184 overexpression and further decreased by miR-184 inhibition in OS cells. In addition, miR-184 agomir reduced doxorubicin-induced cell apoptosis, whereas miR-184 antagomir enhanced apoptosis in OS cells, suggesting that up-regulation of miR-184 contributes to chemoresistance of the OS cell line. CONCLUSIONS Our data show that miR-184 was up-regulated in OS patients treated with doxorubicin therapy and leads to poor response to drug therapy by targeting BCL2L1.
authors.affiliation: Department of Traumatology and Microsurgery, Second People's Hospital of Guangdong Province, The Third Clinical College, Southern Medical University, Guangzhou, Guangdong, China (mainland).
authors.forename: Bo-Chuan
authors.initials: BC
authors.lastname: Lin
authors.affiliation: Department of Traumatology and Microsurgery, Second People's Hospital of Guangdong Province, The Third Clinical College, Southern Medical University, Guangzhou, Guangdong, China (mainland).
authors.forename: Dong
authors.initials: D
authors.lastname: Huang
authors.affiliation: Department of Traumatology and Microsurgery, Second People's Hospital of Guangdong Province, The Third Clinical College, Southern Medical University, Guangzhou, Guangdong, China (mainland).
authors.forename: Chao-Qun
authors.initials: CQ
authors.lastname: Yu
authors.affiliation: Department of Traumatology and Microsurgery, Second People's Hospital of Guangdong Province, The Third Clinical College, Southern Medical University, Guangzhou, Guangdong, China (mainland).
authors.forename: Yong
authors.initials: Y
authors.lastname: Mou
authors.affiliation: Department of Traumatology and Microsurgery,, Second People's Hospital of Guangdong Province, The Third Clinical College, Southern Medical University, Guangzhou, Guangdong, China (mainland).
authors.forename: Yuan-Hang
authors.initials: YH
authors.lastname: Liu
authors.affiliation: Department of Traumatology and Microsurgery, Second People's Hospital of Guangdong Province, The Third Clinical College, Southern Medical University, Guangzhou, Guangdong, China (mainland).
authors.forename: Da-Wei
authors.initials: DW
authors.lastname: Zhang
authors.affiliation: Department of Orthopedics, Daqing Oilfield General Hospital, Daqing, Heilongjiang, China (mainland).
authors.forename: Feng-Jun
authors.initials: FJ
authors.lastname: Shi
mesh_terms: D020413:3' Untranslated Regions; D000903:Antibiotics, Antineoplastic / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects; D001859:Bone Neoplasms / Q000188:drug therapy* / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D004317:Doxorubicin / Q000494:pharmacology*; D019008:Drug Resistance, Neoplasm / Q000235:genetics; D006801:Humans; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D012516:Osteosarcoma / Q000188:drug therapy* / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D051020:bcl-X Protein / Q000235:genetics / Q000378:metabolism*
chemical_list: D020413:3' Untranslated Regions; D000903:Antibiotics, Antineoplastic; C494811:BCL2L1 protein, human; C534424:MIRN184 microRNA, human; D035683:MicroRNAs; D051020:bcl-X Protein; D004317:Doxorubicin