Gaofeng Jiang1, Bo Ren, Lei Xu, Shizhen Song, Changcai Zhu, Fangli Ye. 1. Faculty of Preventive Medicine, Medical College, Huangjiahu Campus, Wuhan University of Science and Technology, Wuhan 430065, PR China. jianggaofeng@yahoo.com.cn
Abstract
BACKGROUND: Survivin, expressed in almost all types of human malignancies, functions as a key factor in radioresistance primarily by inhibiting apoptosis. This study was conducted to investigate whether survivin plays a role in the DNA repair process in the KB human squamous cell carcinoma cell line. MATERIALS AND METHODS: A stable KB cell line overexpressing survivin was established through the use of pIRES2-EGFP vector containing the coding region of survivin. Cells were then irradiated with X-rays and evaluated for DNA double-strand breaks (DSBs) by comet assay and flow cytometry for phospho-histone gammaH2AX. The protein levels of some DSB repair genes were detected by Western blotting analysis. RESULTS: Comet assay and flow cytometry for phospho-histone gammaH2AX showed that overexpression of survivin resulted in significantly fewer DSBs in irradiated cells. Among the DSB repair genes detected, the protein level of Ku70 was up-regulated in survivin-overexpressing KB cells. CONCLUSION: This finding suggests that survivin may enhance DSB repair capability in KB cells by up-regulating Ku70.
BACKGROUND: Survivin, expressed in almost all types of humanmalignancies, functions as a key factor in radioresistance primarily by inhibiting apoptosis. This study was conducted to investigate whether survivin plays a role in the DNA repair process in the KB humansquamous cell carcinoma cell line. MATERIALS AND METHODS: A stable KB cell line overexpressing survivin was established through the use of pIRES2-EGFP vector containing the coding region of survivin. Cells were then irradiated with X-rays and evaluated for DNA double-strand breaks (DSBs) by comet assay and flow cytometry for phospho-histone gammaH2AX. The protein levels of some DSB repair genes were detected by Western blotting analysis. RESULTS:Comet assay and flow cytometry for phospho-histone gammaH2AX showed that overexpression of survivin resulted in significantly fewer DSBs in irradiated cells. Among the DSB repair genes detected, the protein level of Ku70 was up-regulated in survivin-overexpressing KB cells. CONCLUSION: This finding suggests that survivin may enhance DSB repair capability in KB cells by up-regulating Ku70.
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