摘要
目的探讨miR-143在鼻咽癌细胞迁移中的生物学功能及可能的分子机制。方法应用生物信息软件预测miR-143靶基因,分别将预测靶基因GLI3的3'UTR及其突变体构建到荧光素酶载体psiCHECK-2骨架中。通过荧光素酶报告基因检测分析miR-143对靶基因GLI3的调控作用,并检测转染miR-143 mimics、inhibitor和siGLI3后鼻咽癌5-8F细胞miR-143和GLI3的表达水平及迁移能力的变化。结果生物信息学预测Hh信号通路转录基因GLI3可能是miR-143的靶基因;双荧光素酶报告基因分析表明miR-143能够作用于GLI3的3'UTR;Western Blot和qRT-PCR结果进一步证明5-8F细胞中miR-143与GLI3的表达呈负相关,并影响5-8F细胞的迁移能力。结论 MiR-143可通过负调控靶基因GLI3抑制鼻咽癌细胞的迁移,为进一步研究miR-143及Hh信号通路在鼻咽癌中的作用机制提供参考价值。
Objective To investigate the possible biological function and mechanism of miR- 143 in the metastasis of human nasopharyngeal carcinoma （NPC）. Methods Using bioinformatics to predict the target gene of miR-143, the 3＇UTR and mutant 3＇UTR of GLI3 gene was cloned into psiCHECK- 2 vector. Dual- luciferase reporter gene assay was employed to examine the repression of the GLI3 gene. miR-143 and GLI3 expression levels in 5-8F cells transfected with miR-143 mimics, inhibitor, or siGLI3 were examined, and the changes in the cell migration ability was assessed by Transwell invasion assay. Results Bioinformatics prediction indicated the Hh pathway transcription gene GLI3 as a target gene of miR-143, and dual-luciferase reporter assay showed that miR-143 directly combined with the 3’UTR of GLI3. qRT-PCR and Western blotting demonstrated that the expression of miR- 143 in 5- 8F cells was negatively correlated to GLI3 and suppressed the migration of 5- 8F cells. Conclusion MiR-143 can inhibit the invasion of NPC cells by negative regulation of GLI3 gene, which sheds light on the role of miR-143 and Hh pathway in NPC.
出处
《南方医科大学学报》
CAS
CSCD
北大核心
2013年第7期1057-1061,共5页
Journal of Southern Medical University
基金
广东省自然科学基金(S2011040003476)
广东省科技计划项目(2060303)
