探討癌細胞中Eps8於Ku70調控的DNA雙股斷裂修復所扮演的角色

蕭潔君

doi:10.6844/NCKU.2013.00324

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探討癌細胞中Eps8於Ku70調控的DNA雙股斷裂修復所扮演的角色

The Role of Eps8 in Ku70-mediated DNA Double Strand Break Repair in Cancer Cells

蕭潔君 , Masters　　Advisor：呂增宏　　

英文 DOI： 10.6844/NCKU.2013.00324

Eps8 ； Ku70 ； γ-H2AX ； DNA 修復； Eps8 ； Ku70 ； HeLa cell ； DNA damage repairing

Abstract │ Reference (28) │ Altmetrics

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DNA damage, such as base damage and DNA strand breaks, might result in cell apoptosis or cancer formation. Double strand breaks (DSBs) are the primary cytotoxic lesion induced by ionizing radiation (IR) and DNA-targeting anti-cancer drugs. The conventional chemotherapeutic drugs are used together with surgery or radiation therapy to treat metastatic cancers, but usually cause multidrug resistance (MDR). DNA repair ability might be one of the causes of anti-cancer drug resistance. Cancer cells may activate non-homologous end-joining (NHEJ) or homologous recombination (HR) to repair DSBs against DNA-damaging agents. The Ku heterodimer (Ku70/Ku80) is one of the main component of the NHEJ pathway that repairs DNA double-strand breaks (DSBs). We are interested in studying Ku70-interacting proteins, such as Eps8, in DSBs repair. First, we generate HeLa cell expressing Eps8-siRNA or Ku70-siRNA with/without ectopic Eps8. Next, we investigate the participation of Eps8 in the repairing pathway of DNA-damaging agents (doxorubicin and cisplatin) induced DSBs. Single cell electrophoresis (comet assay) is utilized to estimate the degree of DNA damage or DNA repairing ability. We found that doxorubicin- and cisplatin-induced cytotoxicity in Ku70 knockdown cells was decreased by ectopic Eps8. Next, attenuation of Eps8 inhibits DSBs-inudced phosphorylation of H2AX. Intestingly, silencing either Eps8 or Ku70 impaired DSBs repairing. However, ectopically expressed Eps8 further worsen DNA damaging agents-induced DNA damage in Ku70 knockdown cells. This phenomenon can’t be found in cells expressing dominant negative Eps8. In conclusion, Eps8-mediated DSBs repairing might be via a different pathway from Ku70.

Reference ( 28 ) 〈TOP〉

Andrea Kinner., Wenqi Wu., Christian Staudt and George Iliakis. (2008). r-H2AX in recognition and signaling of DNA double-strand breaks in the context of chromatin. Nucleic Acids Research 17, 5678–5694.
連結：
Chen, Y.J., Shen, M.R., Maa, M.C., and Leu, T.H. (2008). Eps8 decreases chemosensitivity and affects survival of cervical cancer patients. Mol Cancer Ther 7, 1376-1385.
連結：
Cheng Q., Barboule N., Frit P., Gomez D., Bombarde O., Couderc B., Ren GS., Salles B., Calsou P.(2011). Ku counteracts mobilization of PARP1 and MRN in chromatin damaged with DNA double-strand breaks. Nucleic Acids Research 22, 9605-9619
連結：
Cohen, H.Y., Lavu, S., Bitterman, K.J., Hekking, B., Imahiyerobo, T.A., Miller, C., Frye, R., Ploegh, H., Kessler, B.M., and Sinclair, D.A. (2004). Acetylation of the C terminus of Ku70 by CBP and PCAF controls Bax-mediated apoptosis. Molecular Cell 13, 627-638.
連結：
Dana Branzei ., Marco Foiani. (2008). Regulation of DNA repair throughout the cell cycle. Molecular cell biology 9, 297-308.
連結：

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