title: Growth-inhibitory effect of neurotrophin-3-secreting adipose tissue-derived mesenchymal stem cells on the D283-MED human medulloblastoma cell line.
abstract: Medulloblastoma (MBL), the most common malignant pediatric brain tumor, is incurable in about one-third of patients and can lead to long-term disabilities despite current multimodal treatments. The purpose of this study was to demonstrate in vitro biological effects of neurotrophins-3 (NT-3) on MBL cells and to evaluate the growth-inhibitory effect of neurotrophin-3 (NT-3)-secreting stem cells on tumor cells. We confirmed by western blotting that D283-MED cells express tyrosine kinase C, a specific receptor for NT-3. Analyzing the biological effects of NT-3 on MBL cells, we evaluated autophagy, apoptosis, senescence, and differentiation of tumor cells with NT-3. The NT-3 induced a concentration-dependent increase in apoptosis in the tumor cell line (P < 0.001). The high concentrations of NT-3 increased the expression of class III β-tubulin (P < 0.001) and decreased the expression of Nestin (P < 0.05). NT-3-secreting stem cells were produced by nucleofecting pIRES2.EGFP-NT3 into human adipose tissue-derived mesenchymal stem cells (hAT-MSCs) and their tropic property toward MBL cells was confirmed by migration assay. Double-layered co-culture experiments with the NT-3-secreting hAT-MSCs and D283-MED MBL cells were performed, and NT-3-induced cell death was studied by 3-(4,5-dimethylathiazol-2-yl)-2,5-dephenyl-tetrazolium bromide (MTT) assay. Consequently, the high concentrations of NT-3-secreting hAT-MSCs significantly (P < 0.05) increased the death of D283-MED cells in vitro. The present study demonstrated that both apoptotic cell death and neuronal differentiation of tumor cells were the mechanisms of growth-inhibitory effect of NT-3-secreting hAT-MSCs on MBL cell line.
authors: Department of Neurosurgery, Seoul National University College of Medicine, Seoul, Korea.
authors: Young-Hoon
authors: YH
authors: Kim
authors: Seung Hee
authors: SH
authors: Cho
authors: Soo Jung
authors: SJ
authors: Lee
authors: Seung Ah
authors: SA
authors: Choi
authors: Ji Hoon
authors: JH
authors: Phi
authors: Seung-Ki
authors: SK
authors: Kim
authors: Kyu-Chang
authors: KC
authors: Wang
authors: Byung-Kyu
authors: BK
authors: Cho
authors: Chae-Yong
authors: CY
authors: Kim
mesh_terms: D000273:Adipose Tissue / Q000166:cytology* / Q000378:metabolism*; D001343:Autophagy; D015153:Blotting, Western; D001932:Brain Neoplasms / Q000473:pathology*; D002454:Cell Differentiation; D002465:Cell Movement; D002469:Cell Separation; D018920:Coculture Techniques; D004396:Coloring Agents; D004797:Enzyme-Linked Immunosorbent Assay; D005455:Fluorescent Antibody Technique; D006801:Humans; D020287:In Situ Nick-End Labeling; D008527:Medulloblastoma / Q000473:pathology*; D059630:Mesenchymal Stem Cells / Q000502:physiology*; D020933:Neurotrophin 3 / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D013778:Tetrazolium Salts; D013844:Thiazoles; D001616:beta-Galactosidase / Q000378:metabolism
chemical_list: D004396:Coloring Agents; D020933:Neurotrophin 3; D013778:Tetrazolium Salts; D013844:Thiazoles; D001616:beta-Galactosidase; C022616:thiazolyl blue