Jiang Lili, Zhang Qingfu, Chang Hongji, Qiu Xueshan, Wang Enhua
Department of Pathology, College of Basic Medical Sciences, China Medical University, Shenyang 110001, China.
Zhongguo Fei Ai Za Zhi. 2009 Oct 20;12(10):1069-73. doi: 10.3779/j.issn.1009-3419.2009.10.03.
MicroRNAs (miRNAs) are endogenous, non-coding small RNA in eukaryotes. They recognize their target sites by incomplete base pairing and posttranscriptionally regulate gene expression, and function on a lot of complex vital processes of organisms. The objective of this work is to study how hsa-miR-125a-5p enhances the invasive ability of lung cancer cells.
The target gene and its target sites of hsa-miR-125a-5p were predicted by microRNA.org. We investigated Rock-1 mRNA and protein expressions by RT-PCR and Western blot according to the result of prediction further. The invasive ability of A549 cells, which were transfected with sense hsa-miR-125a-5p 2'-O-methyl oligonucleotide after being blocked by anti-Rock-1, was observed by Transwell.
With RT-PCR and Western blot, Rock-1 mRNA and protein were both increased in A549 cells transfected with sense hsa-miR-125a-5p 2'-O-methyl oligonucleotide and were both decreased in the cells which transfected with antisense vs control groups. The invasive ability of A549 cells transfected with sense hsa-miR-125a-5p 2'-O-methyl oligonucleotide were weakened after being blocked by anti-Rock-1, vs non-blocking group by Transwell test.
hsa-miR-125a-5p could up-regulate Rock-1 and enhance invasion in lung cancer cells.
微小RNA（miRNA）是真核生物内源性非编码小RNA。它们通过不完全碱基配对识别靶位点，在转录后调控基因表达，并在生物体许多复杂的重要过程中发挥作用。本研究旨在探讨hsa-miR-125a-5p如何增强肺癌细胞的侵袭能力。
通过microRNA.org预测hsa-miR-125a-5p的靶基因及其靶位点。根据预测结果，进一步通过逆转录聚合酶链反应（RT-PCR）和蛋白质免疫印迹法检测Rock-1 mRNA和蛋白表达。采用Transwell法观察抗Rock-1阻断后转染hsa-miR-125a-5p 2'-O-甲基寡核苷酸的A549细胞的侵袭能力。
RT-PCR和蛋白质免疫印迹法检测结果显示，转染hsa-miR-125a-5p 2'-O-甲基寡核苷酸的A549细胞中Rock-1 mRNA和蛋白表达均升高，而转染反义寡核苷酸的细胞中Rock-1 mRNA和蛋白表达均低于对照组。Transwell实验显示，抗Rock-1阻断后，转染hsa-miR-125a-5p 2'-O-甲基寡核苷酸的A549细胞侵袭能力减弱。
hsa-miR-125a-5p可上调Rock-1表达并增强肺癌细胞的侵袭能力。
