Protective Effect of Icariin on DNA Damage Induced by Bleomycin in GC-1 Cells
ZHAO Xueyi, ZHAO Haixia, YANG Yuan, YANG Siqi, YUAN Ding, ZHANG Changcheng*
This study aims to explore the protective effect of ICA (icariin) on BLM (bleomycin)-induced DNA damage in GC-1 cells and its molecular mechanism. GC-1 cells were divided into control group, 10 μg/mL BLM-treated group, BLM+ICA (0.5, 1, 2 and 4 μmol/L) groups. GC-1 cells were pretreated with ICA at different concentrations for 12 h and then added with BLM for another 6 h. Cells were then harvested, and the protein expression levels of DNA damage related proteins γ-H2AX, DNA damage repair related pathway proteins (p-ATM, p-Chk1, p-P53 and P21) and BER (base excision repair)-related pathway proteins (OGG1, APE1 and XRCC1) were detected by Western blot analysis. The expression and localization of γ-H2AX and 8-OHdG were measured by immunofluorescence analysis. Compared with the control group, the protein expression levels of γ-H2AX, p-ATM, p-Chk1, p-P53, P21, OGG1, APE1 and XRCC1 were significantly increased in BLM-treated group. Conversely, ICA diminished the expression levels of γ-H2AX, p-ATM, p-Chk1, p-P53, P21, OGG1, APE1 and XRCC1 in a concentration-dependent manner. The results of immunofluorescence further showed that the number of γ-H2AX positive cells and expression of 8-OHdG were increased in BLM-treated group relative to the control group, whereas, the expression levels of γ-H2AX and 8-OHdG were down-regulated by ICA at different concentrations. In conclusion, ICA attenuates DNA damage induced by BLM in GC-1 cells and down-regulates the ATM/Chk1 and BER signaling pathways.
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