科研之友
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摘要
We investigated microRNAs (miRs) specific to its target gene and exerting distinct biological functions for basal-like breast carcinoma (BLBC). Total RNA was extracted and subjected to miR microarray and bioinformatics analysis. Based on the comprehensive analysis, expression of miRs including its target was analyzed by quantitative reverse transcription-polymerase chain reaction (qRT-PCR), western blot analysis and immunohistochemistry (IHC). Further functional analyses were conducted including proliferation, invasion and apoptosis. miR-205 was identified as down regulated (less than 0.5-fold) in BLBC relatively to normal control (NC). Gene ontology (GO) analysis suggested miR-205 may directly targeted Kruppel-like factor 12 (KLF12; degree=4). Luciferase assay revealed miR-205 directly targeted KLF12 through binding its 3'-untranslated region (3'-UTR; p=0.0016). qRT-PCR and western blot analysis showed miR-205 expression was low in cells (p=0.007) and tumor tissues (n=6; p=0.0074), and KLF12 RNA/protein was observed at high levels in cells (p=0.0026; p=0.0079) and tumor tissues (n=9; p=0.0083); knock-up of miR-205 increased its expression (p=0.0021) but reduced KLF12 RNA/protein levels (p=0.0038; p=0.009) in cells. Modulation of miR-205 expression by transfecting its mimics in cells, was involved in invasion (p=0.00175) and apoptosis (p=0.006). In conclusion, our results supported that miR-205 was a miR specific to BLBC which functioned as tumor suppressor gene through directly targeting and negatively regulating proto-oncogene KLF12. miR-205 dysregulation was involved in invasion and apoptosis. miR-205 and KLF12 provided a potential diagnosis biomarker and therapeutic approach for BLBC.
