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Sci Rep. 2016 Aug 17;6:31534. doi: 10.1038/srep31534.

Recombinant adenovirus of human p66Shc inhibits MCF-7 cell proliferation.

Yang X1,2, Xu R1,2, Lin Y2, Zhen Y3, Wei J2, Hu G2, Sun H1.

Author information

1
Tianjin Key Laboratory of Biomaterial Research, Institute of Biomedical Engineering, Peking Union Medical College &Chinese Academy of Medical Sciences, Tianjin 300192, China.
2
The key Laboratory of Geriatrics, Beijing Hospital &Beijing Institute of Geriatrics, Ministry of Health, Beijing, 100730, China.
3
Department of Histology and Embryology, College of Basic Medical, Hebei United University, Tangshan, 063000, China.

Abstract

The aim of this work was to construct a human recombinant p66Shc adenovirus and to investigate the inhibition of recombinant p66Shc adenovirus on MCF-7 cells. The recombinant adenovirus expression vector was constructed using the Adeno-X Adenoviral System 3. Inhibition of MCF-7 cell proliferation was determined by MTT. Intracellular ROS was measured by DCFH-DA fluorescent probes, and 8-OHdG was detected by ELISA. Cell apoptosis and the cell cycle were assayed by flow cytometry. Western blot were used to observe protein expression. p66Shc expression was upregulated in 4 cell lines after infection. The inhibitory effect of p66Shc recombinant adenovirus on MCF-7 cells was accompanied by enhanced ROS and 8-OHdG. However, no significant differences were observed in the cell apoptosis rate. The ratio of the cell cycle G2/M phase showed a significant increase. Follow-up experiments demonstrated that the expressions of p53, p-p53, cyclin B1 and CDK1 were upregulated with the overexpression of p66Shc. The Adeno-X Adenoviral System 3 can be used to efficiently construct recombinant adenovirus containing p66Shc gene, and the Adeno-X can inhibit the proliferation of MCF-7 cells by inducing cell cycle arrest at the G2/M phase. These results suggested that p66Shc may be a key target for clinical cancer therapy.

PMID:
27530145
PMCID:
PMC4987618
DOI:
10.1038/srep31534
[Indexed for MEDLINE]
Free PMC Article

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