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Mol Med Rep. 2015 Jul;12(1):1107-13. doi: 10.3892/mmr.2015.3549. Epub 2015 Mar 26.

Knockdown of ubiquitin protein ligase E3A affects proliferation and invasion, and induces apoptosis of breast cancer cells through regulation of annexin A2.

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  • 1Department of Anatomy, North Sichuan Medical College, Nanchong, Sichuan 637007, P.R. China.
  • 2Department of Otolaryngology, The Affiliated Hospital of North Sichuan Medical College, Nanchong, Sichuan 637007, P.R. China.
  • 3Morphometric Research Laboratory, North Sichuan Medical College, Nanchong, Sichuan 637007, P.R. China.
  • 4Department of General Surgery, The Affiliated Hospital of North Sichuan Medical College, Nanchong, Sichuan 637007, P.R. China.
  • 5Institute of Immunology and Molecular Biology, North Sichuan Medical College, Nanchong, Sichuan 637007, P.R. China.
  • 6Sichuan Key Laboratory of Medical Imaging, North Sichuan Medical College, Nanchong, Sichuan 637007, P.R. China.

Abstract

The present study used RNA interference (RNAi) to study how the expression of annexin A2 was affected by ubiquitin protein ligase E3A (UBE3A). In addition, the proliferation, apoptosis and invasiveness of BT‑549 breast cancer cells was studied following knockdown of UBE3A. Three pairs of small interfering RNA (siRNA) fragments targeting UBE3A were designed and transfected into the BT‑549 cells. The effects of silencing UBE3A were detected by reverse transcription‑polymerase chain reaction and western blotting, and the same methods were used to detect the expression levels of annexin A2. Cell proliferation was determined using the Cell Counting kit‑8, and flow cytometry and a Transwell chamber assay were used to assess the rate of cell apoptosis and invasion, respectively. Following transfection with the three siRNAs targeting UBE3A for 72 h, the mRNA expression levels of UBE3A were downregulated, as compared with those in the untreated groups, and siRNA1 was the shown to be the most effective siRNA for silencing UBE3A expression. The protein expression levels were concordant with the mRNA expression levels of UBE3A. In addition, the mRNA and protein expression levels of annexin A2 were downregulated. Cellular proliferation and invasion of the siRNA1 group was inhibited as compared with those in the untreated groups, and apoptosis of UBE3A‑siRNA1 cells was increased as compared with that in the untreated groups. The results of the present study indicated that UBE3A may regulate the expression of annexin A2, resulting in promotion of proliferation and invasion and suppression of apoptosis in BT‑549 cells.

PMID:
25816213
[PubMed - in process]
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