FAM92A1-289对人脑胶质瘤U251细胞增殖、迁移能力的影响及其与Galectin-1的关系

发布时间：2018-06-25 22:27

本文选题：脑胶质瘤 + U细胞　；参考：《山东医药》2017年44期

【摘要】：目的探讨FAM92A1-289对人脑胶质瘤U251细胞增殖、迁移能力的影响及其与半乳糖凝集素1(Galectin-1)的关系。方法将CMV-SP6-TALEN-GFP-289质粒转染至U251细胞中,通过嘌呤霉素筛选得到稳定过表达FAM92A1-289基因的脑胶质瘤U251/289++细胞株。取脑胶质瘤U251/289++细胞作为观察组,未转染质粒的野生型U251细胞作为对照组,采用实时无标记动态细胞分析技术检测两组培养24、48、72、96、120 h的细胞增殖能力(以光密度值表示),采用Transwell小室试验检测两组细胞迁移能力(以穿膜细胞数表示)。构建PCS2-3Flag-Galectin-1质粒,将人脑胶质瘤U251细胞随机分为FAM92A1-289组、Galectin-1组、共转染组及空白对照组,FAM92A1-289组、Galectin-1组均采用Lipofactamine 3000转染法分别转染CMV-SP6-TALEN-GFP-289质粒、PCS2-3Flag-Galectin-1质粒,共转染组同时转染上述两种质粒,空白对照组仅加入Lipofactamine 3000转染试剂。采用免疫共沉淀实验验证FAM92A1-289与Galectin-1是否存在相互作用。结果两组培养24 h光密度值比较差异无统计学意义(P0.05);观察组培养48、72、96、120 h光密度值均高于对照组(P0.05或0.01)。观察组与对照组穿膜细胞数分别为(242.0±11.41)、(65.40±7.92)个,两组比较P0.01。共转染组抗GFP蛋白相对表达量明显高于FAM92A1-289组、Galectin-1组及空白对照组(P均0.01)。结论过表达FAM92A1-289可提高人脑胶质瘤U251细胞增殖和迁移能力;FAM92A1-289与Galectin-1之间的相互作用可能为FAM92A1-289调控人脑胶质瘤U251细胞恶性行为的作用机制。
[Abstract]:Objective to investigate the effect of FAM92A1-289 on the proliferation and migration of human glioma U251 cells and the relationship between FAM92A1-289 and galectin-1. Methods the plasmid CMV-SP6-TALEN-GFP-289 was transfected into U251 cells, and the U251 / 289 glioma cell line with stable expression of FAM92A1-289 gene was obtained by purine mycin screening. Brain glioma U251 / 289 cells were used as observation group and wild-type U251 cells with untransfected plasmids as control group. The cell proliferation ability (expressed as optical density) of the two groups was measured by real-time unlabeled dynamic cell analysis technique. The transwell chamber test was used to detect the migration ability of the two groups (expressed by the number of perforated cells). PCS2-3Flag-Galectin-1 plasmid was constructed and human glioma U251 cells were randomly divided into FAM92A1-289 group and FAM92A1-289 group. Both co-transfected group and blank control group (FAM92A1-289) were transfected into CMV-SP6-TALEN-GFP-289 plasmid PCS2-3Flag-Galectin-1 by Lipofactamine 3000. The blank control group only added Lipofactamine 3000 transfection reagent. The interaction between FAM92A1-289 and Galectin-1 was verified by immunoprecipitation. Results there was no significant difference in optical density between the two groups at 24 h (P0.05), while the optical density in the observation group was higher than that in the control group (P0.05 or 0.01), and that in the observation group was higher than that in the control group (P0.05 or 0.01). The number of perforating cells in the observation group and the control group was (242.0 卤11.41), (, 65.40 卤7.92), compared with that in the control group (P 0.01). The relative expression of anti-GFP protein in cotransfection group was significantly higher than that in FAM92A1-289 group and blank control group (P0.01). Conclusion overexpression of FAM92A1-289 can enhance the proliferation and migration of human glioma U251 cells. The interaction between FAM92A1-289 and Galectin-1 may be the mechanism of FAM92A1-289 regulating the malignant behavior of human glioma U251 cells.
【作者单位】：湖北医药学院附属太和医院;湖北医药学院第二临床学院;
【基金】：国家自然科学基金应急管理项目(81641028) 湖北省自然科学基金面上项目(2016CFB408)
【分类号】：R739.41

【相似文献】