MicroRNA-205 directly targets Krüppel-like factor 12 and is involved in invasion and apoptosis in basal-like breast carcinoma

Guan,Bing; Li,Qing; Shen,Li; Rao,Qiu; Wang,Yan; Zhu,Yun; Zhou,Xiao-Jun; Li,Xiao-Hong

doi:10.3892/ijo.2016.3573

MicroRNA-205 directly targets Krüppel-like factor 12 and is involved in invasion and apoptosis in basal-like breast carcinoma

Authors:
- Bing Guan
- Qing Li
- Li Shen
- Qiu Rao
- Yan Wang
- Yun Zhu
- Xiao-Jun Zhou
- Xiao-Hong Li
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Affiliations: Department of Pathology, Shanghai 6th People's Hospital Jinshan Branch, Shanghai 201599, P.R. China, Department of Pathology, Shanghai Pudong New Area People's Hospital, Shanghai 201299, P.R. China, Department of Cardiothoracic Surgery, Shanghai Children's Hospital, Shanghai 201040, P.R. China, Department of Pathology, Nanjing Jinling Hospital, Nanjing, Jiangsu 210002, P.R. China, Department of Pathology, The Second Affiliated Hospital with Nanjing Medical University, Nanjing, Jiangsu 210000, P.R. China, Department of Pathology, Jiangsu Province People's Hospital, Nanjing, Jiangsu 210029, P.R. China
Published online on: June 8, 2016 https://doi.org/10.3892/ijo.2016.3573
Pages: 720-734

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Abstract

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 downregulated (less than 0.5-fold) in BLBC relatively to normal control (NC). Gene ontology (GO) analysis suggested miR-205 may directly targeted Krüppel-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.

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