Journal: British Journal of Cancer
Article Title: Tumour suppressor microRNA-584 directly targets oncogene Rock-1 and decreases invasion ability in human clear cell renal cell carcinoma
doi: 10.1038/sj.bjc.6606028
Figure Lengend Snippet: ROCK-1 is a miR-584 target gene. ( A ) Messenger RNA microarray data of ROCK-1. The miR-584 or miR control were transfected into A-498 cells. At 72 h after transfection, total RNA was extracted and mRNA expression level was compared by Phalanx Bio Inc. ( B ) Sequence of miR-584/ROCK-1 and 3′UTR luciferase assay. The miR-584 binding site in the ROCK-1 3′UTR predicted with microRNA org. (upper). 3′UTR vector and miR-584 or miR control were co-transfected into A-498 cells. Cell lysates were measured for relative luciferase activities at 48 h after transfection. Levels of luciferase activity were compared with those of miR control-transfected cells that is normalised to 1. ( C ) The ROCK-1 protein levels in RCC cells transfected with miR-584. The miR-584 or miR control was transfected into A-498 and 769-P cells. At 72 h after transfection, total protein was extracted and analysed by western blots. β -Actin was used as a loading control. The ratio of band intensity is relative to that of β -actin. Band intensity was measured by using ImageJ software.
Article Snippet: In order to find potential target oncogenes of tumour suppressive miR-584, we used an mRNA microarray service (Phalanx Bio Inc., Palo Alto, CA, USA).
Techniques: Microarray, Control, Transfection, Expressing, Sequencing, Luciferase, Binding Assay, Plasmid Preparation, Activity Assay, Western Blot, Software