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Original Research Article | OPEN ACCESS

Identification of a novel regulatory mechanism involved in inhibition of transcription of suvivin mRNA in breast cancer cells via p21cip-mediated regulation

Hongjie Xu1 , Dajun Wei2, Xiaodong Gai3, Ying Jiang4

1Department of Oncology; 2Department of Cardiology, Affiliated Hospital of Beihua University; 3School of Medical Science, Beihua University, Jilin 132011; 4Health Service Centers of Wenmiao Community, Changyi District, Jilin 132011, Jilin Province, China.

For correspondence:-  Hongjie Xu   Email: xuhongjie0205@sina.com

Accepted: 13 January 2019        Published: 28 February 2019

Citation: Xu H, Wei D, Gai X, Jiang Y. Identification of a novel regulatory mechanism involved in inhibition of transcription of suvivin mRNA in breast cancer cells via p21cip-mediated regulation. Trop J Pharm Res 2019; 18(2):233-241 doi: 10.4314/tjpr.v18i2.3

© 2019 The authors.
This is an Open Access article that uses a funding model which does not charge readers or their institutions for access and distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0) and the Budapest Open Access Initiative (http://www.budapestopenaccessinitiative.org/read), which permit unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited..

Abstract

Purpose: To evaluate the effect of p21Cip1 on survivin transcription levels in breast carcinoma, and to investigate the potential mechanisms.
Methods: Epirubicin, a p21Cip1 activator, was used to treat MCF7 cells. Under the action of normal biological functions of p53, pEGFP-C2-p21 was transfected into MCF7 cells by lipofectamine and positive clones were screened out with G418. The expression levels of p21cip1, p53 and survivin mRNA were quantitated by real-time fluorescent polymerase chain reaction (RQ-PCR). MTT assay was utilized to measure cellular viability and proliferation after transfection. Flow cytometry was employed to determine the cell cycle. Hoechst 33342 staining was carried out to assess cell apoptosis. Lastly, several transcription factor sites located at the promoter region of survivin gene, such as, sp1 site, E2F site and p300/CBP, were measured by p21 overexpression using RT-PCR.
Results: Following epirubicin treatment, within 24 h, the expression levels of endogenous p21cip1 and p53 were up-regulated, whereas that of survivin was down-regulated. After transfection treatment, p21 inhibited the proliferation of MCF7 cells on days 3 and 4, and MCF7 cells overexpressed p21 mRNA, whereas the level of survivin mRNA in MCF7-p21 groups was markedly down-regulated relative to control group, but overexpression of p21 was not sufficient to cause changes in p53 gene expression. The overexpressed p21 resulted in G1/G0 phase arrest based on cell cycle analysis, but apoptosis was not induced. In addition, co-transcription factors E2F-1, sp1 and p300/CBP mRNA levels decreased significantly compared with normal p21 expression groups.
Conclusion: P21cip1 may down-regulate the expression of survivin gene partially by inhibiting the expression level of HAT.

Keywords: Cyclin-dependent kinase inhibitor 1, Phosphoprotein p53, Survivin, Breast carcinoma, G1/G0 phase arrest, Epirubicin, Lipofectamine

Impact Factor
Thompson Reuters (ISI): 0.6 (2023)
H-5 index (Google Scholar): 49 (2023)

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