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目的研究核转运因子CSE1L在化疗后神经母细胞瘤DNA损伤修复过程中的表达变化并初步探索其作用机制,为神经母细胞瘤诊疗提供线索和理论依据。方法通过免疫组化染色方法对19例未化疗和28例经化疗的神经母细胞瘤组织中CSE1L表达水平进行分析。阿霉素处理SH-SY5Y细胞6小时和12小时建立DNA损伤模型,并通过免疫荧光方法检测DNA损伤修复标志物g-H2AX和53BP1在DNA损伤位点的聚集情况;免疫印迹检测阿霉素诱导DNA损伤后CSE1L以及53BP1、FEN-1和EXO-1等DNA损伤相关蛋白的变化水平。慢病毒敲减CSE1L后,免疫印迹检测CSE1L、53BP1、FEN-1和EXO-1蛋白水平变化。结果未化疗组CSE1L阳性率(73.7%)高于化疗组(42.9%)并具有显著差异(卡方检验P=0.037);阿霉素可诱导DNA损伤修复标志物g-H2AX和53BP1在DNA损伤位点的聚集;并诱导53BP1、CSE1L、FEN-1的表达水平在处理6小时先增高、在12小时降低,EXO-1表达持续增高;敲减CSE1L可降低53BP1和FEN-1表达,但促进EXO-1表达。结论 CSE1L在化疗后神经母细胞瘤样本中表达降低,并参与DNA损伤修复过程。
Objective To investigate the expression of CSE1L in the process of DNA damage repair of neuroblastoma after chemotherapy, and to explore its mechanism of action, providing clues and theoretical basis for the diagnosis and treatment of neuroblastoma. Methods The expression levels of CSE1L in 19 cases of non-chemotherapy and 28 cases of chemotherapy-treated neuroblastoma were analyzed by immunohistochemical staining. Doxorubicin treatment SH-SY5Y cells at 6 hours and 12 hours to establish a DNA damage model, and by immunofluorescence detection of DNA damage repair markers g-H2AX and 53BP1 at DNA damage site aggregation; Western blot detection of doxorubicin induction Changes of CSE1L, DNA damage related proteins such as 53BP1, FEN-1 and EXO-1 after DNA damage. After CSE1L was knocked down by lentivirus, the protein levels of CSE1L, 53BP1, FEN-1 and EXO-1 were detected by Western blotting. Results The positive rate of CSE1L in non-chemotherapy group (73.7%) was significantly higher than that in chemotherapy group (42.9%) (P = 0.037). Adriamycin induced DNA damage repair markers g-H2AX and 53BP1 in DNA damage The expression of 53BP1, CSE1L and FEN-1 were increased at 6 hours and decreased at 12 hours, while EXO-1 expression continued to increase. Knockdown of CSE1L decreased the expression of 53BP1 and FEN-1, EXO-1 expression. Conclusion The expression of CSE1L in the neuroblastoma samples after chemotherapy is decreased and involved in DNA damage repair process.