Effect of Capn4 in repairing DNA damage of nasopharyngeal carcinoma cell CNE2
WANG Hui, WANG Feng, WU Li-xian, ZHENG Ming
Chinese Journal of Clinical Anatomy ›› 2017, Vol. 35 ›› Issue (3) : 291-295.
Effect of Capn4 in repairing DNA damage of nasopharyngeal carcinoma cell CNE2
Objective To investigate the effect of Capn4 on the repair of nasopharyngeal carcinoma cell line CNE2 induced by radiation and its possible pathways. Methods The protein levels of Capn4 in CNE2 and 293T cells were examined by Western blot. The Capn4 knockdown cells using shRNA was constructed in CNE2 cells and further identified by Western blot. Highly Connected Cell Imaging Analyzer was used to analyze the Mean Fluorescence Intensity of γ-H2AX in CNE2 cells after radiation and CNE2 cells with down-regulation of Capn4. Non-homologous end joining (NHEJ) pathway-specific plasmids EJ5-GFP were transfected into CNE2 cells with Fugen6 HD and CNE2 cells with downregulation of Capn4. The positive rate of GFP was detected by Highly Connected Cell Imaging Analyzer. The downregulation of Canp4 on NHEJ repair pathway of DNA damage was induced by radiation influences. Results The expression levels of Capn4 were significantly higher in CNE2 cells than in 293T cells. The shRNA inhibited Capn4 expression in CNE2 cells. Downregulation of Capn4 expression could inhibit the DNA damage of CNE2 cells induced by radiation（P<0.01）. Downregulation of Capn4 expression could inhibit the repair of NHEJ pathway in CNE2 cells after DNA damage（P<0.01）. Conclusion Capn4 is highly expressed in nasopharyngeal carcinoma CNE2 cells. Downregulation of Capn4 expression can inhibit radiation-induced DNA damage repair, which may be achieved by inhibition of NHEJ pathway.
Capn4； Nasopharyngeal carcinoma； / Radiation； / shRNA； / NHEJ pathway
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