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目的:探讨miR-92a调节RECK表达对髓母细胞瘤细胞(Daoy)增殖迁移的影响并研究其可能的分子机制。方法:脂质体将miR-92a前体和阴性对照转染Daoy细胞,重组腺病毒介导RECK在Daoy细胞的表达,采用Taqman试剂盒检测miR-92a的表达,Real time PCR检测转染后细胞中RECK等基因的表达,Western blot检测细胞中RECK等蛋白的表达,MTT检测细胞增殖能力,Transwell检测细胞侵袭迁移能力。结果:Daoy细胞中miR-92a表达高于miR-17~92基因簇的其他成员。过表达miR-92a增加Daoy细胞的增殖和迁移能力;miR-92a的过表达抑制RECK的蛋白表达;而腺病毒介导的RECK的过表达,则可以抑制Daoy细胞的增殖和迁移。结论:miR-92a可能通过降低RECK的表达来提高Daoy细胞的增殖和迁移能力。
AIM: To investigate the effect of miR-92a regulating RECK expression on proliferation and migration of medulloblastoma cells (Daoy) and to investigate its possible molecular mechanism. METHODS: Daoy cells were transfected with miR-92a precursor and negative control by liposome. The expression of RECK in Daoy cells was induced by recombinant adenovirus. The expression of miR-92a was detected by Taqman kit. The expression of miR-92a was detected by Real time PCR. RECK and other genes expression, Western blot detection of RECK protein expression in cells, MTT assay of cell proliferation, Transwell assay of cell invasion and migration. Results: The expression of miR-92a in Daoy cells was higher than other members of miR-17-92 gene cluster. Overexpression of miR-92a increased the proliferation and migration of Daoy cells; overexpression of miR-92a inhibited the expression of RECK protein; and adenovirus-mediated RECK overexpression inhibited the proliferation and migration of Daoy cells. Conclusion: miR-92a may enhance the proliferation and migration of Daoy cells by decreasing the expression of RECK.