Abstract:DAXX was initially identified as a pro-apoptotic protein that bound to the death domain of the CD95 death receptor. By activating the c-Jun NH2-terminal kinasepathway, DAXX was showed to enhance both CD95-mediated and transforming growth factor-β-dependent apoptosis. In this study, we constructed a tetracycline-inducible (tet-on) DAXX silencing lentiviral vector, packaged lentiviral particles, then infected OV2008 cells, cells were selected and cultured in complete cell culture medium along with either puromycin (1μg/mL) for 7~10 days. DAXX was efficiently knocked down in these cells with tetracycline. DAXX protein and gene expression were detected by Western blotting and Q-PCR, cells proliferation and clone number of DAXX-depleted OV2008 cells were detected by MTT assay and colony forming assay. DNA damage protein expression of DAXX-depleted cells was detected by Western blotting and immunofluorescence. Senescent cells were detected by SA-β-galactosidase staining. The results showed that tetracycline induced dose-dependent decrease of DAXX. DAXX depletion significantly inhibited the proliferation and colony forming of OV2008 cells, as assessed by MTT proliferation assay and colony forming assay. Western blotting results showed that p-H2AX and p-CHK2 expressions were significantly increased after DAXX silencing. DAXX silencing induced senescence were determined by SA-β-gal staining. Western blotting results showed that p21 and p27 expressions were significantly increased after DAXX silencing. The results showed that DAXX silencing inhibited ovarian cancer cell proliferation, colony forming, and promoted DNA damage insults and senescence. Our results give new idea in the field of gene therapy for ovarian cancer.
潘巍巍 曹利仙 沈忠飞 徐营. 沉默DAXX基因表达促进人卵巢癌细胞凋亡和衰老[J]. 中国生物化学与分子生物学报, 2014, 30(7): 691-699.
PAN Wei-Wei CAO Li-Xian SHEN Zhong-Fei XU Ying. Silencing DAXX Gene Promotes Apoptosis and Senescence in Cultured Ovarian Cancer Cells. Chinese Journal of Biochemistry and Molecular Biol, 2014, 30(7): 691-699.
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