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过表达SNORD47对U87-EGFRvIII胶质瘤细胞增殖和侵袭的影响

Effect of SNORD47 over-expression on proliferation and invasion of U87-epidermal growthfactor receptorvIII glioma cells
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摘要 目的探讨过表达SNORD47对U87-EGFRvIII胶质瘤细胞增殖和侵袭的影响。方法将对数生长期U87垣GFRvIII胶质瘤细胞分为SNORD47组、阴性对照组、空白对照组3组，前2组分别转染携带SNORD47核苷酸序列、阴性对照核苷酸序列的重组慢病毒，空白对照组不作任何处理。48h后qRT—PCR检测3组细胞SNORD47的表达水平：Westernblotting检测3组细胞基质金属蛋白酶2（MMP2）、MMP9蛋白的表达；CCK-8实验检测接种后4、24、48、72、96h3组细胞的存活率：平板克隆实验检测3组细胞的克隆能力；划痕实验、Transwell侵袭实验分别检测3组细胞的迁移能力和侵袭能力。结果qRT-PCR和Westernblotting检测显示：与阴性对照组及空白对照组比较，SNORD47组细胞SNORD47表达水平升高，MMP2、MMP9蛋白表达水平降低，差异均有统计学意义（P〈0．05）；CCK-8和平板克隆实验显示：与阴性对照组及空白对照组比较，SNORD47组细胞接种后48、72、96h吸光度值降低，SNORD47组细胞克隆数降低，差异均有统计学意义（P〈0．05）：划痕实验、Transwell侵袭实验显示：与阴性对照组及空白对照组比较，SNORD47组划痕损伤面积增大，穿膜细胞数明显减少，差异有统计学意义（P〈0．05）。结论SNORD47能有效抑制u87-EGFRvIII胶质瘤细胞的增殖和侵袭能力。 Objective To study the effect of SNORD47 over-expression on proliferation and invasion of U87-epidermal growth factor receptor （EGFR）vIII glioma cells. Methods U87-EGFRvIII glioma cells at logarithmic phase were assigned into lenti-SNORD47 group, lenti-NC group and blank control group. The recombinant lentiviruses containing lenti-SNORD47 or lenti-NC were transfected into U87-EGFRvlII glioma cells of the lenti-SNORD47 group and lenti-NC group, respectively. Forty-eight h after transfection, the SNORD47 expression in the three groups was measured by real time quantitative PCR. Western blotting was used to detect the protein expressions of matrix metalloproteinase （MMP） 2 and MMP9. The proliferation of U87-EGFRvIII cells 4, 24, 48, 72 and 96 h after transfection was evaluated by CCK-8 assay and colony formation assay. Transwell assay and wound-healing assay were used to examine the invasion and migration of these cells. Results The SNORD47 expression in the lenti-SNORD47 group was significantly higher than that in the lenti-NC group and control group （P〈0.05）. The protein expressions of MMP2 and MMP9 in the lenti-SNORD47 group were significantlydecreased as compared with those in the lenti-NC group and control group （P〈0.05）. At 48, 72 and 96 h after transfection, the optical density and number of cloned cells in the lenti-SNORD47 group were significantly decreased as with those in the lenti-NC group and control group （/＇〈0.05）. The invasion and migration abilities of U87-EGFRvlII cells in the lenti-SNORD47 group were significantly suppressed as compared with those in the lenti-NC group and control group （P〈0.05）. Conclusion SNORD47 could inhibit the proliferative and invasive abilities ofU87-EGFRvIII glioma cells.
作者 徐斌 叶敏华 吕世刚 吴淼经 肖兵 范阳华 柴毅 祝新根
机构地区 南昌大学第二附属医院神经外科
出处 《中华神经医学杂志》 CSCD 北大核心 2017年第6期547-552,共6页 Chinese Journal of Neuromedicine
基金 国家自然科学基金（81660420） 江西省优势科技创新团队建设计划（20152BCB24009） 江西省对外科技合作计划（20151BDH80009）
关键词 SNORD47 神经胶质瘤 细胞增殖 细胞侵袭 细胞迁移 基质金属蛋白酶 SNORD47 Glioma Proliferation Invasion Migration Matrixmetalloproteinase
分类号 R739.41 [医药卫生—肿瘤]
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中华神经医学杂志
2017年 第6期

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