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

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

摘要

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

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.

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