Glucose transporter protein 1-targeted RNA interference inhibits growth and invasion of the osteosarcoma cell line MG63 in vitro.

Cancer biotherapy & radiopharmaceuticals Pub Date : 2010-10-01 Epub Date: 2010-09-21 DOI:10.1089/cbr.2010.0784
Jian Fan, Jia-Qian Zhou, Guang-Rong Yu, Dong-Dong Lu
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引用次数: 18

Abstract

Malignant cells show increased glucose uptake, which is thought to be mediated by glucose transporters. Glucose transporter protein 1 (Glut-1) is critical for growth, proliferation, and migration of tumor cells and Glut-1 overexpression is associated with poor overall survival in osteosarcoma patients. The present study was designed to determine the role of Glut-1 in the growth and invasion of the osteosarcoma cell line MG63, using RNA interference technology in vitro. shRNA-expressing lentiviral vectors targeting the Glut-1 gene were constructed, which were stably expressed in MG63 cells. The level of Glut-1 mRNA was investigated using real-time reverse transcription-polymerase chain reaction, and the protein expression of Glut-1 mRNA was observed using western blotting. MG63 cellular glucose uptake, proliferation, and migration were detected by methyl thiazole tetrazolium assay and flow cytometry. A Glut-1-specific shRNA-expressing lentiviral vector was obtained, which could efficiently inhibit the mRNA and protein expression of Glut-1 to ∼82%-85% in MG63 cells. Downregulation of Glut-1 inhibited the cellular glucose uptake, growth, and invasion of MG63 cells in vitro. These results indicate that RNA interference targeting of Glut-1 could be an effective strategy for the treatment of osteoscarcoma patients.

葡萄糖转运蛋白1靶向RNA干扰对骨肉瘤MG63细胞生长和侵袭的体外抑制作用
恶性细胞表现出葡萄糖摄取增加,这被认为是由葡萄糖转运蛋白介导的。葡萄糖转运蛋白1 (Glut-1)对肿瘤细胞的生长、增殖和迁移至关重要,Glut-1过表达与骨肉瘤患者的总生存率低有关。本研究旨在利用RNA干扰技术,确定Glut-1在骨肉瘤MG63细胞系生长和侵袭中的作用。构建了靶向Glut-1基因的shrna慢病毒载体,并在MG63细胞中稳定表达。实时逆转录-聚合酶链反应检测Glut-1 mRNA水平,western blotting检测Glut-1 mRNA蛋白表达。采用甲基噻唑四唑实验和流式细胞术检测MG63细胞葡萄糖摄取、增殖和迁移。获得了一种表达谷氨酸-1特异性shrna的慢病毒载体,该载体能有效抑制MG63细胞中谷氨酸-1 mRNA和蛋白的表达至82%-85%。Glut-1的下调抑制了MG63细胞对葡萄糖的摄取、生长和侵袭。这些结果表明,靶向Glut-1的RNA干扰可能是治疗骨肉瘤患者的有效策略。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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