title: Recombinant adenovirus of human p66Shc inhibits MCF-7 cell proliferation.
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.
authors.affiliation: Tianjin Key Laboratory of Biomaterial Research, Institute of Biomedical Engineering, Peking Union Medical College &Chinese Academy of Medical Sciences, Tianjin 300192, China.|The key Laboratory of Geriatrics, Beijing Hospital &Beijing Institute of Geriatrics, Ministry of Health, Beijing, 100730, China.
authors.forename: Xiaoshan
authors.initials: X
authors.lastname: Yang
authors.affiliation: Tianjin Key Laboratory of Biomaterial Research, Institute of Biomedical Engineering, Peking Union Medical College &Chinese Academy of Medical Sciences, Tianjin 300192, China.|The key Laboratory of Geriatrics, Beijing Hospital &Beijing Institute of Geriatrics, Ministry of Health, Beijing, 100730, China.
authors.forename: Rong
authors.initials: R
authors.lastname: Xu
authors.affiliation: The key Laboratory of Geriatrics, Beijing Hospital &Beijing Institute of Geriatrics, Ministry of Health, Beijing, 100730, China.
authors.forename: Yajun
authors.initials: Y
authors.lastname: Lin
authors.affiliation: Department of Histology and Embryology, College of Basic Medical, Hebei United University, Tangshan, 063000, China.
authors.forename: Yongzhan
authors.initials: Y
authors.lastname: Zhen
authors.affiliation: The key Laboratory of Geriatrics, Beijing Hospital &Beijing Institute of Geriatrics, Ministry of Health, Beijing, 100730, China.
authors.forename: Jie
authors.initials: J
authors.lastname: Wei
authors.affiliation: The key Laboratory of Geriatrics, Beijing Hospital &Beijing Institute of Geriatrics, Ministry of Health, Beijing, 100730, China.
authors.forename: Gang
authors.initials: G
authors.lastname: Hu
authors.affiliation: Tianjin Key Laboratory of Biomaterial Research, Institute of Biomedical Engineering, Peking Union Medical College &Chinese Academy of Medical Sciences, Tianjin 300192, China.
authors.forename: Hongfan
authors.initials: H
authors.lastname: Sun
mesh_terms: D000080242:8-Hydroxy-2'-Deoxyguanosine; D000256:Adenoviridae / Q000235:genetics* / Q000502:physiology; D059447:Cell Cycle Checkpoints / Q000235:genetics; D049109:Cell Proliferation / Q000502:physiology*; D003849:Deoxyguanosine / Q000031:analogs & derivatives / Q000378:metabolism; D057809:HEK293 Cells; D006367:HeLa Cells; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D061986:MCF-7 Cells; D010766:Phosphorylation; D017382:Reactive Oxygen Species / Q000378:metabolism; D011995:Recombination, Genetic; D000071425:Src Homology 2 Domain-Containing, Transforming Protein 1 / Q000235:genetics* / Q000378:metabolism; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism; D015854:Up-Regulation
chemical_list: D017382:Reactive Oxygen Species; C528024:SHC1 protein, human; D000071425:Src Homology 2 Domain-Containing, Transforming Protein 1; C495901:TP53 protein, human; D016159:Tumor Suppressor Protein p53; D000080242:8-Hydroxy-2'-Deoxyguanosine; D003849:Deoxyguanosine