title: [Down-regulation of miR-146b-5p promotes malignant transformation of fusion cells after co-culture of macrophages with glioma stem cells in vitro].
abstract: Objective:To observe mutual interactions between macrophages(Mφ) and glioma stem cells (GSCs)in dual-color tracing model in vitro, to identify the biological characteristics of fusion cells in multiple levels, and to analysis the relevant molecular mechanisms.Methods:Red fluorescent protein(RFP) gene was stably transfected into human GSCs cell line SU4. Mφ cells were obtained from Balb/c nude mice with enhanced green fluorescent protein (EGFP) expression. Then two cells were co-cultured in dual-color tracing platform. RFP/EGFP double positive cells with high proliferation ability were mono-cloned. The fusion cells were verified by Western blot, fluorescence in situ hybridization, immunocytochemistry and chromosome karyotype analysis.The biological characteristics of fusion cells were further analyzed, together with relevant molecular changes.Results:RFP / EGFP double positive cells were obtained through in vitro co-culture. RFP and EGFP coexpression were proved at transcriptional and translational levels in the fusion cells. They also co-expressed GSCs marker Nestin and Mφ marker CD68, and karyotype analysis showed two types of characteristic chromosomes, which confirmed that the fusion cells originated from spontaneous fusion between SU4-RFP and Mφ.Fusion cell proliferation rate and invasion ability were higher than SU4-RFP, which were relevant with down-regulation of miR-146b-5p and activation of STAT3. Fusion cells transfected with miR-146b-5p showed a higher apoptosis rate(18.83%) and lower tumor formation(4/5).Conclusion:Mφ could fuse with GSCs spontaneously in local tumor micro-environment. The proliferation and invasion abilities of fusion cells were higher than their parent cells, which were relevant with down-regulation of miR-146b-5p and activation of STAT3. It revealed the possible mechanisms of malignant progression of gliomas.
authors.affiliation: Department of Neurosurgery, the Second Affiliated Hospital of Soochow University, Suzhou 215004, China.
authors.forename: H H
authors.initials: HH
authors.lastname: Cai
authors.forename: H Y
authors.initials: HY
authors.lastname: Wang
authors.forename: H R
authors.initials: HR
authors.lastname: Liu
authors.forename: Y J
authors.initials: YJ
authors.lastname: Sheng
authors.forename: D G
authors.initials: DG
authors.lastname: Xi
authors.forename: Y P
authors.initials: YP
authors.lastname: Xue
authors.forename: X L
authors.initials: XL
authors.lastname: Dai
authors.forename: A D
authors.initials: AD
authors.lastname: Wang
authors.forename: Q
authors.initials: Q
authors.lastname: Huang
authors.forename: J
authors.initials: J
authors.lastname: Dong
mesh_terms: D000818:Animals; D002450:Cell Communication; D002459:Cell Fusion; D045744:Cell Line, Tumor; D002471:Cell Transformation, Neoplastic; D018920:Coculture Techniques; D015536:Down-Regulation; D005910:Glioma; D049452:Green Fluorescent Proteins; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D008164:Luminescent Proteins; D008264:Macrophages; D051379:Mice; D008819:Mice, Nude; D035683:MicroRNAs; D014411:Neoplastic Stem Cells; D014162:Transfection; D000097573:Red Fluorescent Protein
chemical_list: D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins; D035683:MicroRNAs; D000097573:Red Fluorescent Protein; C485184:enhanced green fluorescent protein
keywords: Dual-color fluorescent tracing; Glioma stem cells; Macrophages; Malignant transformation; miR-146b-5p