PTEN mRNA编辑的间充质干细胞对神经胶质瘤U251细胞杀伤作用研究

doi:10.13560/j.cnki.biotech.bull.1985.2016.04.032

摘要/Abstract

摘要： 间充质干细胞（Mesenchymal stem cells，MSCs）具有向肿瘤组织迁移趋化的特性，所以它被认为是一种携带特殊基因进行肿瘤的靶向治疗的理想载体。与不同形式的DNA载体相比，体外合成的mRNA更容易转染并且不会引入突变。利用间接共培养方法研究PTEN（gene of phosphate and tension homology deleted on chromsome ten，PTEN）mRNA编辑的MSCs对U251-Luc细胞杀伤作用。体外转录合成PTEN mRNA，转染到MSCs，利用Western blot方法检测转染后PTEN蛋白表达情况，transwell共培养方法分析转染PTEN mRNA对MSCs肿瘤细胞迁移趋化性影响。利用活体成像仪和荧光显微镜检测在间接共培养条件下PTEN mRNA编辑的MSCs对U251-Luc细胞杀伤作用。体外成功合成PTEN mRNA，转染到U251-Luc细胞后能正确表达PTEN蛋白，并且在转染24 h表达最高。与对照组相比，转染PTEN mRNA后能一定程度提高MSCs细胞对肿瘤迁移能力（P<0.05）。PTEN mRNA编辑的MSCs培养上清能显著抑制U251-Luc细胞生长（P<0.05）。体外合成抑癌基因mRNA的方法为MSC介导的靶向基因治疗神经胶质瘤提供新思路。

关键词: 间充质干细胞, 合成mRNA, PTEN, 基因治疗, U251神经胶质瘤细胞

Abstract: Mesenchymal stem cells（MSCs）have been considered of great potential as ideal carriers for the delivery of anticancer agents since the discovery of their capacity of tumor-oriented migration and integration. Differing from DNA-based vectors，synthesized mRNA in vitro owns the properties of its easy transfection and mutagenesis-free. This study aims to investigate the effects of phosphatase and tensin homolog（PTEN）mRNA-engineered MSCs on human glioma U251 cells under indirect co-culture condition. PTEN mRNA by in vitro transcription was transfected into MSCs，and then the expression of protein PTEN in the transfected MSCs was detected by Western blot，and the migration and integration effects of PTEN mRNA transfection on MSC tumor cells were verified using transwell co-cultures. The cytotoxic effects of PTEN mRNA-engineered MSCs on the U251-Luc glioma cells were detected with luminescence and fluorescence microscopy under indirect co-culture condition. The in vitro synthesized PTEN mRNA was transfected to U251-Luc cells，protein PTEN expressed correctly and the expression was the highest at 24 hours after transfection. An enhanced migration rate was observed with MSCs transfected with PTEN mRNA compared to non-transfected MSCs（P<0.05）. A significant inhibition of U251 cells was observed when they were cultured with conditioned medium from PTEN mRNA-engineered MSCs（P<0.05）. The results suggest that anticancer gene-bearing mRNA synthesized in vitro provides new insights into the MSC-mediated targeted gene therapy of glioblastoma.

Key words: mesenchymal stem cells, synthesized mRNA, PTEN, gene therapy, U251 glioma cell

郭兴荣,王小莉,袁雅红,涂汉军. PTEN mRNA编辑的间充质干细胞对神经胶质瘤U251细胞杀伤作用研究[J]. 生物技术通报, 2016, 32(4): 234-241.

GUO Xing-rong, WANG Xiao-li, YUAN Ya-hong ,TU Han-jun. The Cytotoxic Effects of PTEN-mRNA-engineered Mesenchymal Stem Cell on U251 Glioma Cells[J]. Biotechnology Bulletin, 2016, 32(4): 234-241.

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