Notch1 regulates stemness and chemotherapeutic sensitivity of human glioma U251 cells |
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Authors: | ZHANG Li-ke XIAN Na LIN Ling GONG Yu-qing YE Zhi-qiang ZHENG Zhi-hong |
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Affiliation: | Research Center of Neurobiology, Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Fujian Medical University, Fuzhou 350108, China |
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Abstract: | AIM: To investigate whether Notch1 changes stemness and chemotherapeutic sensitivity in human glioma U251 cells. METHODS: The lentiviral vectors, which expressed Notch1-shRNA or Notch1 intracellular domain (NICD), were transfected into U251 cells. Western blot and immunofluorescence staining were applied to monitor the validity of the cells, down-regulation of Notch1 expression or over-expression of NICD. The proportion of CD133+ cells was analyzed by flow cytometry. The expression of nestin and GFAP was identified by immunofluorescence staining. The formation rate of tumor cell spheres and the implanted tumor growth in SCID mice were observed. MTT assay was performed to evaluate the chemotherapeutic sensitivity to VM-26 and BCNU of the cells with different treatments. RESULTS: Stemness was significantly enhanced in the cells over-expressing NICD. For example, the proportion of CD133+ cells was increased, the expression of nestin was up-regulated, the expression of GFAP was down-regulated, and the formation rate of tumor cell spheres and implanted tumor growth were increased. The chemotherapeutic sensitivity to VM-26 and BCNU of the cells was decreased. In the cells with Notch1 gene down-regulation by RNAi, the stemness was inhibited and chemotherapeutic sensitivity was increased. CONCLUSION: Notch1, which leads to the change of stemness and chemotherapeutic sensitivity in human glioma U251 cells, is likely to be a potential molecular target for treatment of glioma. |
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Keywords: | Glioma U251 cells Notch1 Chemotherapeutic sensitivity |
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