【摘要】 目的阐明MALAT1通过miR-30对骨巨细胞瘤基质细胞生长、迁移和侵袭进行调控。方法收集临床骨巨细胞瘤及对照样本进行荧光定量PCR,检测MALAT1及miR-30表达水平;对骨巨细胞瘤进行原代细胞培养,获得纯化的骨巨细胞瘤基质细胞原代细胞系;构建MALAT1过表达质粒,合成miR-30mimic类似物及inhibitor抑制剂;在细胞系中过表达MALAT1质粒和/或miR-30相关片段,通过MTT实验观察细胞生长情况,通过Transwell实验观察细胞迁移和侵袭情况。结果与正常组织对比,MALAT1表达水平在骨巨细胞瘤组织中显著降低,而miR-30水平升高;生物信息学分析发现MALAT1存在于miR-30相互作用的序列,且荧光素酶报道基因结果显示,过表达miR-30可显著抑制MALAT1 WT质粒的报道基因信号,而对MUT突变质粒无效;单独过表达MALAT1或者miR-30 inhibitor抑制剂均可抑制基质细胞生长、迁移和侵袭;而且上下游拮抗实验表明,过表达MALAT1可抵消miR-30 mimic类似物引起的促进作用。结论 MALAT1、miR-30均参与骨巨细胞瘤基质细胞发展调控;MALAT1可通过调控miR-30抑制骨巨细胞瘤细胞增殖、迁移和侵袭。更多还原

【Abstract】 Objective To elucidate whether MALAT1 regulates the growth,migration,and invasion of giant cell tumor stromal cells through miR-30.Methods Collect the samples of cancer and para-cancer in clinical patients with giant cell tumor of bone for fluorescence quantitative PCR,to detect the expression level of MALAT1 and miR-30;culture primary cells of giant cell tumor of bone to obtain purified primary stromal cell line;construct MALAT1 overexpression plasmid,synthesize miR-30 mimic and inhibitor;overexpress MALAT1 plasmid and/or miR-30 related fragments in cells,observe cell growth by MTT experiment,observe migration and invasion by transwell experiment.Results Compared with the adjacent tissues,the expression level of MALAT1 was significantly decreased in giant cell tumor of bone tissue,while the level of miR-30 was increased;bioinformatics analysis found that MALAT1 showed matched sequence interacting with miR-30,and the luciferase reporter gene reporter showed that overexpression of miR-30 can significantly inhibit the luciferase signal of MALAT1 WT,but not for MUT mutant plasmids;overexpression of MALAT1 or miR-30 inhibitor alone inhibited the growth,migration,and invasion of stromal cells;and downstream antagonism experiments showed that overexpression of MALAT1 counteracted the promotion effect induced by miR-30 mimic.Conclusion MALAT1 and miR-30 are involved in the development and regulation of bone giant cell tumor stromal cells;MALAT1 inhibits the proliferation,migration,and invasion of bone giant cell tumor cells by regulating miR-30.更多还原