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Published on: April 6, 2012
MicroRNA let-7a inhibits proliferation of breast cancer cell by downregulating USP32 expression
Chunyan Liu1, Zhaobo Chen2, Min Fang3
1Department of Integrated Traditional Chinese and Western Medicine, Medical College of Qingdao University, Qingdao 266021, China.
Background:
The present study aimed to investigate the effect of microRNA (miR) let-7a on ubiquitin specific protease 32 (USP32) expression and its potential function in MCF-7 breast cancer (BCa) cell line.
Methods:
BCa MCF-7 cells were transfected with hsa-miR let-7a mimics or inhibitors, then the USP32 expression was evaluated using quantitative reverse transcription polymerase chain reaction (qRT-PCR) and western blot analysis in the transfected cells. USP32 as a target regulated by miR let-7a was confirmed via Dual-luciferase reporter assay. The effects of miR let-7a on the viability were determined using MTT assay and colony formation analysis.
Results:
Western blot analysis revealed that miR let-7a mimics dramatically decreased the USP32 protein expression, whereas miR let-7a inhibitors increased the protein expression of USP32 compared with their controls in the MCF-7 cells. Dual-luciferase reporter assay showed that miR let-7a mimics could directly target the 3'-untranslated region (UTR) of USP32. Further, MTT assay and colony formation analysis showed that miR let-7a significantly inhibited cell proliferation of MCF-7 cells. However, overexpression of USP32 could reverse the effect of miR let-7a on MCF-7 cells proliferation.
Conclusions:
Collectively, the results suggested that miR let-7a functions as a tumor suppressor to reduce proliferation by targeting USP32 in BCa cells.
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