摘要
目的预测和验证miR-339-5p下游直接靶基因。方法利用生物信息学方法预测miR-339-5p潜在靶基因;克隆目标基因3’UTR,将其与含荧光素酶报告基因的质粒psiCHECK-2连接后,与miR-339-5p mimics共转染乳腺癌细胞株T47D,做荧光素酶报告实验验证miR-339-5p靶基因;人乳腺癌细胞株T47D转染miR-339-5p mimics(模拟物)后,行Western blot法检测靶基因蛋白表达。结果生物信息学方法预测结果显示BCL-6为miR-339-5p潜在靶基因之一;荧光素酶报告实验显示,miR-339-5p可直接调节BCL-6基因3’UTR;Western blot法检测结果显示T47D细胞转染miR-339-5p mimics后,BCL-6蛋白表达水平明显下调。结论 miR-339-5p可以调节下游靶基因BCL-6的表达。
Purpose To predict and verify the direct target gene of miR-339-Sp in breast cancer. Methods Bioinformatics tools were used to analysis and predict the potential target genes of miR-339-Sp. The 3'UTR region of target gene was cloned to the psiCHECK-2 vector which including luciferase reporter gene and then co-trausfected with miR-339-Sp mimics to T47D. MiR-339-Sp target genes were confirmed by luciferase reporter assay. The protein expression of taget gene was detected by transfection of miR-339-5p mimics to T47D cell though western-blot. Results BCL-6 is one of the potential target genes of miR-339-Sp by bioinformatics tools analysis. The luciferase reporter assay indicated that miR-339-Sp was directly bound to BCL-6 3 ' UTR. The expression level of BCL-6 protein was significantly decreased in T47D cell with forced expression of miR-339-Sp. Conclusion MiR-339-Sp can regulate the expression of downstream target gene BC-6 in breast cancer cell.
出处
《临床与实验病理学杂志》
CAS
CSCD
北大核心
2013年第3期244-246,共3页
Chinese Journal of Clinical and Experimental Pathology
基金
国家自然科学基金资助(81101597
81172533)