摘要
目的微小RNA(microRNA)是一类长度约20 bp的非编码RNA,在多种疾病包括肿瘤的发生发展中发挥重要作用。miR-451在多种恶性肿瘤中呈低表达,并且与肿瘤发生发展密切相关。已有研究发现miR-451在骨肉瘤组织中表达显著降低,且miR-451低表达提示患者预后较差,总生存期和无瘤生存期较短。但是,miR-451促进骨肉瘤发生发展的分子机制尚未阐明。方法本研究检测了miR-451过表达对骨肉瘤细胞侵袭和上皮间质转化(EMT)的作用。我们首先构建miR-451过表达的慢病毒载体,然后将慢病毒转染U2OS细胞以提高miR-451的表达水平;采用划痕试验、Transwell实验观察miR-451过表达对骨肉瘤细胞迁移和侵袭的影响;采用Western blot实验检测miR-451过表达对上皮间质转化相关蛋白E-cadherin、Vimentin以及转录因子NF-κB活性的影响。结果 miR-451过表达在体外可以显著抑制U2OS细胞迁移和侵袭,可以上调U2OS细胞内E-cadherin蛋白表达而下调Vimentin蛋白表达,并且可以显著下调NF-κB关键亚单位IKK-β蛋白水平。结论 miR-451可能通过调控NF-κB活性从而发挥其抑制骨肉瘤细胞迁移、侵袭和EMT作用。
Objective MicroRNA is a class of small non-coding RNAs containing about 22 nucleotides, which has been implicated in many disease including tumors, miR-451 has been shown to be downregulated in several human malignan- cies and correlated with tumor progression. It has been demonstrated that miR-451 was significantly decreased in osteosar- coma tissues, and the patients with low miR-451 expression have shorter overall and disease-free survival. However, the underlying molecular mechanisms by which miR-451 contributes to osteosareoma tumorigenesis have not been elucidated yet. Methods In the present study, the effect of miR-451 overexpression on invasion and epithelial-mesenchymal transi- tion （EMT） of osteosarcoma was investigated. First, the lentivirus carrying miR-451 mimics were constructed and trans- fected into U2OS cells to elevate miR-451 expression. Wound healing test and Transwell assay were conducted to examine the effects of miR-451 overexpression on the migration and invasion of U2OS cells in vitro. Western blot assay was carried out to examine the EMT-associated proteins such as E-cadherin, Vimentin and transcription factor NF-KB. Results The miR-451 overexpression suppressed cell migration and invasion, up-regulated the expression of E-cadherin, down-regula- ted the expression of Vimentin, and down-regulated the expression of the expression of IKK-β. Conclusion The miR- 451 may suppress the migration, invasion and EMT through inhibiting the activities of NF-κB.
作者
陈晓东
邓敏
CHEN Xiao-dong;DENG Min(Department of Orthopedics, the First Affiliated Hospital of Bengbu Medical College, Bengbu, Anhui 233004, Chin)
出处
《中华全科医学》
2018年第7期1064-1066,1226,共4页
Chinese Journal of General Practice
基金
安徽省自然科学基金一般项目(KJ2015B042by)
蚌埠医学院科技发展基金项目(BYKF1737)
蚌埠医学院院级课题(Byky1340)
安徽省高校省级青年人才基金重点项目(2013SQRL053ZD)
