capture of microrna–bound mrnas identifies the tumor suppressor mir-34a as a regulator of growth factor signaling捕获microrna-bound mrna识别肿瘤抑制mir-34a作为生长因子信号调节器.pdfVIP

Capture of MicroRNA–Bound mRNAs Identifies the Tumor Suppressor miR-34a as a Regulator of Growth Factor Signaling Ashish Lal1,2,3.*, Marshall P. Thomas1,2., Gabriel Altschuler4., Francisco Navarro1,2., Elizabeth O’Day1,2, 3 5 5 1,2 1,2 Xiao Ling Li , Carla Concepcion , Yoon-Chi Han , Jerome Thiery , Danielle K. Rajani , Aaron 1,2 4 5 4 1,2 Deutsch , Oliver Hofmann , Andrea Ventura , Winston Hide , Judy Lieberman * 1 Immune Disease Institute, Program in Cellular and Molecular Medicine, Children’s Hospital Boston, Boston, Massachusetts, United States of America, 2 Department of Pediatrics, Harvard Medical School, Boston, Massachusetts, United States of America, 3 Genetics Branch, National Cancer Institute, National Institutes of Health, Bethesda, Maryland, United States of America, 4 Department of Biostatistics, Harvard School of Public Health, Boston, Massachusetts, United States of America, 5 Department of Cancer Biology and Genetics, Memorial Sloan Kettering Cancer Center, New York, New York, United States of America Abstract A simple biochemical method to isolate mRNAs pulled down with a transfected, biotinylated microRNA was used to identify direct target genes of miR-34a, a tumor suppressor gene. The method reidentified most of the known miR-34a regulated genes expressed in K562 and HCT116 cancer cell lines. Transcripts for 982 genes were enriched in the pull-down with miR- 34a in both cell lines. Despite this large number, validation experiments suggested that ,90% of the genes identified in both cell lines can be directly regulated by miR-34a. Thus miR-34a