https://scholars.lib.ntu.edu.tw/handle/123456789/594005
標題: | Neural differentiation of glioblastoma cell lines via a herpes simplex virus thymidine kinase/ganciclovir system driven by a glial fibrillary acidic protein promoter | 作者: | Luo, Elizabeth Wei-Chia MENG-LIN LIAO CHUNG-LIANG CHIEN |
關鍵字: | CANCER STEM-CELLS; ASTROCYTE-SPECIFIC EXPRESSION; SUICIDE GENE-THERAPY; IDENTIFICATION; CYTOTOXICITY; HSVTK; BRAIN | 公開日期: | 2021 | 出版社: | PUBLIC LIBRARY SCIENCE | 卷: | 16 | 期: | 8 | 來源出版物: | PloS one | 摘要: | Glioblastoma is a malignant brain tumor with poor prognosis that rapidly acquires resistance to available clinical treatments. The herpes simplex virus thymidine kinase/ganciclovir (HSVtk/GCV) system produces the selective elimination of HSVtk-positive cells and is a candidate for preclinical testing against glioblastoma via its ability to regulate proliferation and differentiation. Therefore, in this study, we aimed to establish a plasmid encoding the HSVtk/GCV system driven by a glial fibrillary acidic protein (GFAP) promoter and verify its possibility of neural differentiation of glioblastoma cell line under the GCV challenge. Four stable clones-N2A-pCMV-HSVtk, N2A-pGFAP-HSVtk, U251-pCMV-HSVtk, and U251-pGFAP-HSVtk-were established from neuronal N2A and glioblastoma U251 cell lines. In vitro GCV sensitivity was assessed by MTT assay for monitoring time- and dosage-dependent cytotoxicity. The capability for neural differentiation in stable glioblastoma clones during GCV treatment was assessed by performing immunocytochemistry for nestin, GFAP, and βIII-tubulin. Under GFAP promoter control, the U251 stable clone exhibited GCV sensitivity, while the neuronal N2A clones were nonreactive. During GCV treatment, cells underwent apoptosis on day 3 and dying cells were identified after day 5. Nestin was increasingly expressed in surviving cells, indicating that the population of neural stem-like cells was enriched. Lower levels of GFAP expression were detected in surviving cells. Furthermore, βIII-tubulin-positive neuron-like cells were identified after GCV treatment. This study established pGFAP-HSVtk-P2A-EGFP plasmids that successfully ablated GFAP-positive glioblastoma cells, but left neuronal N2A cells intact. These data suggest that the neural differentiation of glioblastoma cells can be promoted by treatment with the HSVtk/GCV system. |
URI: | https://scholars.lib.ntu.edu.tw/handle/123456789/594005 | ISSN: | 1932-6203 | DOI: | 10.1371/journal.pone.0253008 |
顯示於: | 解剖學暨細胞生物學科所 |
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