Targeting c-Myc on cell growth and vascular endothelial growth factor expression in IN500 glioblastoma cells
Targeting c-Myc on cell growth and vascular endothelial growth factor expression in IN500 glioblastoma cells
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靶向 c-Myc 对 IN500 胶质母细胞瘤细胞生长和血管内皮生长因子表达的影响
DOI：
10.3760/cma.j.issn.0366-6999.2012.11.033
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发表时间：
2012-06-05
影响因子：
6.1
通讯作者：
Chen Ze
中科院分区：
文献类型：
--
作者：
Hu Yu-hua;Kong Shi-qi;Chen Ze
研究背景c-Myc表达水平与胶质瘤病理分级高、预后差密切相关。血管内皮生长因子（VEGF）是最重要的血管生成因子，能有效刺激血管内皮细胞的增殖和迁移。本研究旨在阐明c-Myc在胶质瘤发生发展中的生物学意义，我们下调了c-Myc在人胶质母细胞瘤细胞系IN 500中的表达，并研究了c-Myc在体外对细胞生长、增殖、方法用shRNA稳定转染IN 500 Delta细胞，观察转染后细胞凋亡和VEGF表达的变化，以及对肿瘤形成的影响。表达c-Myc的质粒（pCMYC-shRNA）或作为对照的质粒（pCtrl-shRNA）。建立稳定的细胞系后，采用逆转录（RT）-PCR检测c-Myc和VEGF mRNA的表达，采用Western blotting和免疫组化检测c-Myc和VEGF蛋白的表达。流式细胞仪检测细胞周期进程和凋亡。结果稳定转染pCMYC-shRNA后，IN 500细胞中c-Myc的mRNA和蛋白表达水平均明显下降，且与VEGF表达下调呈正相关。在体外下调c-Myc也导致G1-S阻滞和增强的细胞凋亡。在体内，靶向c-Myc减少异种移植瘤的形成，并导致显着更小的tumors.Conclusions c-Myc在胶质母细胞瘤的发展，包括调节细胞周期，凋亡和VEGF的表达具有多种功能。靶向c-Myc表达可能是治疗恶性胶质瘤的一种有希望的方法。中华医学杂志2012;125（11）：2025-2031
Background The level of c-Myc is closely associated with high pathological grade and the poor prognosis of gliomas. Vascular endothelial growth factor (VEGF) is the most important angiogenic factor that potently stimulates the proliferation and migration of vascular endothelial cells. This study aimed to address the biological importance of c-Myc in the development of gliomas, we downregulated the expression of c-Myc in the human glioblastoma cell line IN500 and studied the in vitro effect on cellular growth, proliferation, and apoptosis and the expression of VEGF and the in vivo effect on tumor formation in a xenograft mouse model.Methods IN500 Delta cells were stably transfected with shRNA-expressing plasmids for either c-Myc (pCMYC-shRNA) or as a control (pCtrl-shRNA). Following establishment of stable cells, the mRNA expressions of c-Myc and VEGF were examined by reverse transcription (RT)-PCR, and c-Myc and VEGF proteins by Western blotting and immunohistochemistry. Cell-cycle progression and apoptosis were determined by flow cytometry. The in vivo effect of targeting c-Myc was determined by subcutaneous injection of stable cells into immunodeficient nude mice.Results The stable transfection of pCMYC-shRNA successfully knocked down the steady-state mRNA and protein levels of c-Myc in IN500, which positively correlated with the downregulation of VEGF. Downregulating c-Myc in vitro also led to G1-S arrest and enhanced apoptosis. In vivo, targeting c-Myc reduced xenograft tumor formation and resulted in significantly smaller tumors.Conclusions c-Myc has multiple functions in glioblastoma development that include regulating cell-cycle, apoptosis, and VEGF expression. Targeting c-Myc expression may be a promising therapy for malignant glioma. Chin Med J 2012;125(11):2025-2031