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[Reversal of resistance to adriamycin in human breast cancer cell line MCF-7/ADM by silencing AEG-1 gene and its mechanism.] [Article in Chinese]

YUAN Lei, SHI Ran-Ran, RAO Shu-Mei, SONG Jin-ling, CUI Ming-Chen^*

Laboratory of Molecular Biology, Luohe Medical College, Luohe 462002, China

Abstract

The aim of this study was to investigate the effects of AEG-1 gene silencing on the chemoresistance of human breast cancer cell line MCF-7/ADM and its possible mechanism. MCF-7/ADM cells were incubated in the medium containing adriamycin (ADM). The recombinant pLKO.1-shAEG-1 plasmid was constructed to silence AEG-1 expression in human breast cancer MCF-7/ADM cells. MTT assay was employed to detect the anti-tumor effect of ADM on MCF-7/ADM cells, and IC50 value of ADM was calculated according to MTT. Flow cytometry was used to determine the apoptosis. Western blot was used to analyze the expression levels of AEG-1, p-Akt, p-MDM2, p-Bad, p53 and MDR1. The result showed MCF-7/ADM had a significantly higher expression level of AEG-1 compared with that of MCF-7 (P < 0.05), however, the expression of AEG-1 was decreased after AEG-1 gene silencing. The IC50 value of ADM in shAEG-1 group was significantly lower than that in shcontrol group. AEG-1 gene silencing induced cell apoptosis and enhanced the pro-apoptotic effect of ADM on MCF-7/ADM cells. After AEG-1 gene silencing, the phosphorylation of Akt, MDM2 and Bad was inhibited (P < 0.05), the protein levels of p53 and MDR1 were up-regulated (P < 0.05) and down-regulated (P < 0.05) respectively, compared with control. In conclusion, the results suggest that AEG-1 gene silencing can reverse the ADM resistance in human breast cancer cell line MCF-7/ADM by means of inducing apoptosis and down-regulating the protein level of MDR1.

Key words: astrocyte elevated gene-1; MCF-7/ADM; apoptosis; multidrug resistance gene 1; Akt

Received: 2014-03-07　　Accepted: 2014-04-17

Corresponding author: 崔明辰　　E-mail: cuimingchen123@126.com

Citing This Article：

YUAN Lei, SHI Ran-Ran, RAO Shu-Mei, SONG Jin-ling, CUI Ming-Chen. [Reversal of resistance to adriamycin in human breast cancer cell line MCF-7/ADM by silencing AEG-1 gene and its mechanism.] [Article in Chinese]. Acta Physiol Sin 2014; 66 (5): 625-630 (in Chinese with English abstract).