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.
Weiderpass E, Labrèche F.Malignant tumors of the female reproductive system[J].Saf Health Work, 2012, 3(3):166-180
Wong AS, Leung PC.Role of endocrine and growth factors on the ovarian surface epithelium[J].J Obstet Gynaecol Res, 2007, 33(1):3-16
Jemal A, Siegel R, Ward E, et al.Cancer statistics2009[J].CA Cancer J Clin, 2009, 59(4):225-249
Wethkamp N, Hanenberg H, Funke S, et al.Daxx-beta and Daxx-gamma,two novel splice variants of the transcriptional co-repressor Daxx[J].J Biol Chem, 2011, 286(22):19576-19588
Yun HJ, Yoon JH, Lee JK, et al.Daxx mediates activation-induced cell death in microglia by triggering MST1 signalling[J].EMBO J, 2011, 30(12):2465-2476
Michaelson JS, Leder P.RNAi reveals anti-apoptotic and transcriptionally repressive activities of DAXX[J].J Cell Sci, 2003, 116(Pt 2):345-352
Morozov VM, Massoll NA, Vladimirova OV, et al.Regulation of c-met expression by transcription repressor Daxx[J].Oncogene, 2008, 27(15):2177-2186
Muromoto R.Death domain-associated protein (DAXX)-mediated regulation of transcription and cell death[J].Yakugaku Zasshi, 2012, 132(9):979-984
AbediniM R, Muller E J, Bergeron R, et al.Akt promotes chemoresistance in human ovarian cancer cells by modulating cisplatin-induced,p53-dependent ubiquitination of FLICE-like inhibitory protein[J].Oncogene, 2010, 29(1):11-25
Pennington K, Pulaski H, Pennington M, et al.Too much of a good thing: suicide prevention promotes chemoresistance in ovarian carcinoma[J].Curr Cancer Drug Targets, 2010, 10(6):575-583
Zhang B, Pan J S, Liu J Y, et al.Effects of chemotherapy orand radiotherapy on survivin expression in ovarian cancer[J].Methods Find Exp Clin Pharmacol, 2006, 28(9):619-625
Heaphy CM, de Wilde RF, Jiao Y, et al.Altered telomeres in tumors with ATRX and DAXX mutations[J].Science, 2011, 333(6041):425-
Jiao Y, Shi C, Edil BH, et al.DAXXATRX,MEN1,and mTOR pathway genes are frequently altered in pancreatic neuroendocrine tumors[J].Science, 2011, 331(6021):1199-
Chaudhry P, Srinivasan R, Patel FD.Expression of the major fas family and Bcl-2 family of proteins in epithelial ovarian cancer (EOC) and their correlation to chemotherapeutic response and outcome[J].Oncol Res, 2010, 18(11-12):549-559
Giovinazzi S, Lindsay CR, Morozov VM, et al.Regulation of mitosis and taxane response by Daxx and Rassf1[J].Oncogene, 2012, 31(1):13-26
Bellovin DI, Das B, Felsher DW.Tumor dormancy, oncogene addiction, cellular senescence, and self-renewal programs [J]. Adv Exp Med Biol, 2013, 734: 91-107
Courtois-Cox S, Genther Williams SM, Reczek EE, et al.A negative feedback signaling network underlies oncogene-induced senescence[J].Cancer Cell, 2006, 10(6):459-472
Dankort D, Filenova E, Collado M, et al.A new mouse model to explore the initiation,progression,and therapy of BRAFV600E-induced lung tumors[J].Genes Dev, 2007, 21(4):379-384
Sarkisian CJ, Keister BA, Stairs DB, et al.Dose-dependent oncogene-induced senescence in vivo and its evasion during mammary tumorigenesis[J].Nat Cell Biol, 2007, 9(5):493-505
Campisi J, d'Adda di Fagagna F.Cellular senescence: when bad things happen to good cells[J].Nat Rev Mol Cell Biol, 2007, 8(9):729-740
