Fig. 5

Leukemia inhibitory factor receptor (LIFR) is a direct target of miR-629-3p. a The putative sequence of the LIFR 3′-untranslated region bound to the seed region of miR-629-3p. b The luciferase activity of wild-type LIFR reporter constructs in MDA-MB-231 cells exhibited a distinct reduction after cotransfection with miR-629-3p compared with those cotransfected with scrambled oligonucleotides. However, mutant LIFR abrogated the posttranscriptional inhibitory effect of miR-629-3p. c Quantitative real-time polymerase chain reaction analysis indicated that transfection of MCF-7 cells with miR-629 or LIFR short-hairpin RNA interfering plasmid (shLIFR) led to a significant reduction in LIFR messenger RNA (mRNA) levels. Meanwhile, transfection with anti-miR-629 or LIFR-expressing lentiviral plasmid (Lv-LIFR) increased the expression of LIFR in MDA-MB-231 cells compared with the corresponding negative controls. d Western blot analysis showed that transfection of MCF-7 cells with miR-629, shLIFR, or shLIFR + miR-629 led to a significant reduction in LIFR protein levels (P < 0.05), especially in the shLIFR + miR-629 group (P < 0.01). Meanwhile, transfection with anti-miR-629, Lv-LIFR, or Lv-LIFR + anti-miR-629 increased the expression of LIFR in MDA-MB-231 cells significantly (P < 0.05), especially in the Lv-LIFR + anti-miR-629 group (P < 0.01). The inhibitory effect of miR-629 and shLIFR on LIFR expression was similar and also promoted the effect of anti-miR-629 and Lv-LIFR. * P < 0.05 and ** P < 0.01