The effect of downregulation of Stathmin gene on biological behaviors of U373 and U87-MG glioblastoma cells

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.

Association of STMN1 with survival in solid tumors: A systematic review and meta-analysis

“…37 Furthermore, some researchers demonstrated that the downregulation of STMN1 expression was capable of inhibiting the migration and invasion of cancer cells. 14,52,53,56 Overall, these previous studies support the notion that STMN1 may act as an oncogene and plays a pivotal role in promoting cancer progression, indicating that STMN1 may be a useful prognostic biomarker and therapeutic target. Consistently, accumulating evidence has suggested that elevated STMN1 expression is closely correlated not only with the development and progression of cancer, 10,11 but also with poor survival.…”

“…[49][50][51] Similarly, STMN1 downregulation could also inhibit the proliferation of glioblastoma cells and esophageal squamous cell carcinoma. 14,52 Additionally, several previous studies revealed that silencing the expression of STMN1 promoted cellular apoptosis of glioma, 50 oral cancer, 53 prostate cancer, 54 non-small cell lung cancer, 55 and esophageal squamous cell carcinoma. 37 Furthermore, some researchers demonstrated that the downregulation of STMN1 expression was capable of inhibiting the migration and invasion of cancer cells.…”

Oleanolic acid mediated the proliferation and invasion of U251 glioma cells and promoted their apoptosis through the IKK-β, MAPK3, and MAPK4 signaling pathway

“…In addition to the poor therapeutic effect of chemotherapy, early radiotherapy, and late adjuvant therapy on malignant glioma, these treatments also have toxic side effects. Therefore, identifying new therapeutic agents to improve the prognosis of patients with malignant glioma is critical [10,11] .…”

Oleanolic Acid Inhibits the Proliferation and Invasion of U251Glioma Cells and Promotes their Apoptosis through the IKK-β, MAPK3, and MAPK4 Signaling Pathway

“…In addition to the poor therapeutic effect of chemotherapy, early radiotherapy, and late adjuvant therapy on malignant glioma, these treatments also have toxic side effects. Therefore, identifying new therapeutic agents to improve the prognosis of patients with malignant glioma is critical [12,13] . To overcome the limitations of the existing treatment methods for malignant glioma, many new treatment methods such as molecular targeted therapy, immunotherapy, gene therapy, stem cell therapy, and nanotechnology have begun to appear in preclinical and clinical research.…”