干扰MAP2基因表达对胶质瘤细胞生物学特性的影响

目的:通过RNA干扰降低胶质瘤细胞中MAP2表达后观察其对胶质瘤细胞生物学特性的影响。方法:RT-qPCR和Western blot测定胶质瘤细胞Hs683、U251中MAP2的基因和蛋白表达。构建shRNA MAP2质粒，转染胶质瘤细胞，筛选出MAP2低表达的胶质瘤细胞，并经Western blot验证。对转染后胶质瘤细胞行MTT检测细胞活性、PI-FACS检测细胞周期、Annexin V-APC单染法行细胞周期检测、Transwell实验检测细胞的迁移能力，并使用Western blot测定转染后胶质瘤细胞中α-管蛋白和乙酰化α-管蛋白的含量。结果:转染shRNA MAP2后胶质瘤细胞中MAP2的蛋白表达显著降低。分子生物学实验显示转染shRNA MAP2后胶质瘤细胞的活性降低、凋亡增加、细胞迁移能力下降、细胞周期有所变化但无特异性，代表微管稳固性的乙酰化α-管蛋白含量基本不变。结论:RNA干扰降低胶质瘤细胞Hs683、U251中MAP2表达后，能降低细胞活性、增加凋亡、降低细胞迁移能力，但这些变化似与微管的稳固性无关。

Effect of silencing MAP2 on the biological characteristics of glioma cells

Objective:In this study,MAP2 expression in glioma cells was reduced using RNA interference.Furtherly,the effects on the biological characteristics of glioma cells was observed.Methods:Real-time quantitative polymerase chain reaction and Western blot were employed to determine the expression of MAP2 in glioma cell lines Hs683 and U251.A stable expression plasmid with shRNA MAP2 was constructed and transfected into glioma cell lines.The shRNA MAP2 glioma cells with stable and low expression were selected and confirmed by Western blot analysis.MTT assay was used to detect the cell viability of each group of glioma cells.Annexin V-APC single staining method was used to detect the cell apotosis.PI-FACS was used to detect cell cycle distribution.Transwell test was used to detect the activity of cell migration,and Western blot was used to detect the protein expression of α-tubulin and acetylated α-tubulin.Results:The expression of MAP2 protein in shRNA MAP2 glioma cells was significantly lower than the control groups.After transfection with shRNA MAP2,compared with control cell lines,the cell viability and migrative ability were decreased.The cell apoptosis was increased.The cell cycle distribution was changed,but not specifically,and the expression of acetylated α-tubulin,representative of microtubule stability,were neither changed.Conclusion:RNA interference silencing MAP2 gene in glioma cells can decrease cell viability and migrative ability,increase cell apoptosis,change the cell cycle distribution.But these effects may have no relation with microtubule stability.