Abstract
BackgroundStathmin as a critical protein involved in microtubule polymerization, is necessary for survival of cancer cells. However, extremely little is known about Stathmin in glioblastoma. So, this study was designed to elucidate the function of Stathmin gene in the tumorigenesis and progression of glioblastoma cells.MethodThe lentiviral interference vector pLV3-si-Stathmin targeting Stathmin gene and the control vector pLV3-NC were established for the co-transfection of 293T cells together with the helper plasmids. Viral titer was determined via limiting dilution assay. Then pLV3-si-Stathmin and pLV3-NC were stably co-transfected into U373 and U87-MG glioblastoma cells. Expression levels of Stathmin protein in each group were determined by using Western Blot, and the proliferation and migration ability of the cells with downregulated Stathmin were evaluated through CCK8 assay and transwell invasion assay, respectively. Cell cycles and cell apoptosis were detected with flow cytometry. Finally, the effect of Stathmin in tumor formation was determined in nude mice.ResultDNA sequencing and viral titer assay indicated that the lentiviral interference vector was successfully established with a viral titer of 4 × 108 TU/ml. According to the results from Western Blotting, Stathmin protein expression level decreased significantly in the U373 and U87-MG cells after transfected with pLV3-si-Stathmin, respectively, compared with those transfected with pLV3-NC. In glioblastoma cells, the cell proliferation and migration were greatly inhibited after the downregulation of Stathmin protein. Flow cytometry showed that much more cells were arrested in G2/M phasein Stathmin downregulated group, compared with the non-transfection group and NC group. But Stathmin downregulation did not induce significant cell apoptosis. Tumor formation assay in nude mice showed that tumor formation was delayed after Stathmin downregulation, with a reduction in both tumor formation rate and tumor growth velocity.ConclusionStathmin downregulation affected the biological behaviors of U373 and U87-MG glioblastoma cells, inhibiting the proliferation and migration of tumor cells. Stathmin gene may serve as a potential target in gene therapy for glioblastoma.
Highlights
Stathmin as a critical protein involved in microtubule polymerization, is necessary for survival of cancer cells
Stathmin downregulation affected the biological behaviors of U373 and U87-MG glioblastoma cells, inhibiting the proliferation and migration of tumor cells
Stathmin gene may serve as a potential target in gene therapy for glioblastoma
Summary
Stathmin as a critical protein involved in microtubule polymerization, is necessary for survival of cancer cells. This study was designed to elucidate the function of Stathmin gene in the tumorigenesis and progression of glioblastoma cells. Stathmin is involved in the assembly and regulation of microtubules and spindles through binding to the tubulin. It participates in cell proliferation, differentiation, regeneration and motion via binding to different proteins [3]. We demonstrated that downregulation of Stathmin expression inhibited the cell proliferation, cell migration and tumor formation in nude mice of U373 and U87-MG glioblastoma cells. Stathmin affects glioblastoma progression through regulating cell proliferation, cell cycle and cell migration. Our finding demonstrates that Stathmin gene might serve as a potential therapeutic target in glioblastoma
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