miR-137调控人脑胶质瘤细胞增殖侵袭能力的体内外研究

目的探讨miR-137调控人脑胶质瘤细胞的增殖侵袭性生长能力及其机制。方法采用实时PCR分析miR-137在不同品系胶质瘤细胞及不同级别胶质瘤样本中的表达;脂质体介导miR-137模拟物转染胶质瘤细胞,实时PCR检测转染后miR-137的表达;应用MTT法、流式细胞术评价细胞生长和增殖的生物学特征变化;划痕实验、transwell细胞体外迁移实验检测肿瘤细胞迁移、侵袭能力;动物实验评价体内条件下肿瘤生长能力变化;Western blot、免疫组织化学染色检测肿瘤细胞Ki-67、MMP9表达水平。结果实时PCR分析显示：miR-137在胶质瘤中低表达。miR-137模拟物转染LN229和U87细胞后,实时PCR显示：miR-137表达上调;MTT法及流式细胞术显示细胞生长受抑,出现G0/G1期阻滞;划痕实验及transwell实验证实：细胞迁移侵袭能力下降;进一步Western blot、免疫组织化学染色显示：增殖侵袭相关蛋白Ki-67、MMP9表达降低;动物实验反映：肿瘤细胞生长受抑制。结论 miR-137高表达可抑制胶质瘤细胞生长和侵袭能力,提示miR-137可作为基因治疗脑胶质瘤的候选靶点。

In vitro and vivo research of proliferation and invasion potentials of human glioma cells regulated by miR-137

Objective To study the potentials and mechanism of proliferation and invasion of human glioma cells regulated by miR-137. Methods Real-time PCR was used to identify the expression of miR-137 in different cells. The miR-137 mimics were transfected into human glioma cells, then the expression of miR-137 was detected by real-time PCR. The changes of cell growth and proliferation were determined by MTT method and flow cytometry assay. The migration and invasion ofglioma cells were detected by wound scratch assay and Transwell assay. The change of tumor growth in vivo was evalUated by animal experiment. The expressions of Ki-67 and MMP9 were detected by Western blotting and immunohistochemical staining. Results Low expression of miR-137 was found in glioma cells by real-time PCR, and miR-137 was up-regulated after the LN229 and U87 cells transfection with miR-137 mimics. The proliferation was significantly decreased by the MTT assay and the cell cycle was arrested in G0/G1 phase by flow cytometry assay after transfection with miR-137 mimics. Scratch assay and Transwell assay demonstrated that the migration and invasion capability of tumor cells decreased after transfection with miR-137 mimics, and decreased expressions of Ki-67 and MMP9 were detected by Western blotting and immunohistochemical staining. The animal experiment showed that cell growth was inhibited. Conclusion High expression of miR-137 can inhibit the proliferation and invasion of human glioma cells, which suggests that miR-137 can be considered as a.candidate target for gene treatment of glioma.