沉默ski基因对大鼠星形胶质细胞活化增殖及Cyclin D1表达的影响

发布时间：2019-01-05 05:28

【摘要】：目的研究ski基因对大鼠星形胶质细胞增殖的影响及其分子机制。方法分离3日龄Sprague-Dawley大鼠脑皮质星形胶质细胞,体外培养。合成ski和阴性对照小干扰片段。设置ski-si RNA组、阴性对照组和空白对照组,采用脂质体法将特异性针对ski基因的si RNA和阴性对照si RNA转入ski-si RNA组、阴性对照组星形胶质细胞中。Western blotting分析ski、胶质纤维酸性蛋白(GFAP)和Cyclin D1蛋白表达;CCK8法检测星形胶质细胞增殖活力;流式细胞仪检测细胞周期。结果 ski-si RNA组星形胶质细胞ski蛋白表达明显降低(F=38.611,P0.01),GFAP(F=7.547,P0.05)和Cyclin D1蛋白(F=3.901,P0.05)表达降低;培养12 h后,ski-si RNA组增殖活力明显降低(F30.507,P0.01),G1期细胞比例明显增加,S期明显减少(F48.425,P0.01)。结论沉默ski基因可能通过下调Cyclin D1表达,抑制星形胶质细胞增殖。
[Abstract]:Objective to study the effect of ski gene on astrocyte proliferation and its molecular mechanism. Methods the cortical astrocytes of 3 day old Sprague-Dawley rats were isolated and cultured in vitro. The small interference fragments of ski and negative control were synthesized. In ski-si RNA group, negative control group and blank control group, si RNA specific for ski gene and negative control si RNA were transferred into ski-si RNA group by liposome method. Ski, in astrocytes of negative control group was analyzed by. Western blotting. The expression of glial fibrillary acidic protein (GFAP) (GFAP) and Cyclin D1 protein; The proliferative activity of astrocytes was detected by CCK8 and the cell cycle was detected by flow cytometry. Results the expression of ski protein and Cyclin D1 protein in astrocytes of ski-si RNA group were significantly decreased (P 0.05) and Cyclin D1 protein (FN 3.901P 0.05). After 12 hours of culture, the proliferation activity of ski-si RNA group was significantly decreased (F30.507, P0.01), the proportion of cells in G1 phase was obviously increased, and that in S phase was significantly decreased (F48.425, P0.01). Conclusion silencing ski gene may inhibit the proliferation of astrocytes by down-regulating the expression of Cyclin D1.
【作者单位】：兰州大学第二医院骨科;甘肃省骨关节疾病研究重点实验室;
【基金】：国家自然科学基金项目(No.30772299) 兰州大学第二医院科研基金项目(No.sdkyjj-04)
【分类号】：R3416

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