中文摘要：

目的：探讨赖氨酰氧化酶（LOX）和XRCC5在人乳腺癌细胞内是否存在相互作用。方法：构建含有LOX全长及插入LOX基因序列C端的2个StrepⅡ和一个Flag标签的慢病毒表达载体GV303/EGFP（LOX-SF）,经酶切鉴定正确后,将其转染MDA-MB-231人乳腺癌细胞,使用荧光定量PCR和蛋白质印迹实验对细胞中重组蛋白LOX-SF进行检测。利用免疫共沉淀技术验证LOX与XRCC5之间的相互作用,并通过间接免疫荧关分析LOX与XRCC5在MDA-MB-231人乳腺癌细胞的定位情况。结果：重组LOX蛋白在MDA-MB-231人乳腺癌细胞中稳定表达。LOX抗体免疫共沉淀下来的蛋白复合物中检测到XRCC5,同时XRCC5抗体免疫共沉淀下来的蛋白复合物中也检测到LOX。利用间接免疫荧光分析,LOX与XRCC5在细胞核内存在共定位。结论：LOX与XRCC5在人乳腺癌细胞内存在相互作用,并且在细胞核内共定位。

英文摘要：

Objective：To investigate possible interaction between LOX and XRCC5 in human breast cancer cell. Methods：The full-length LOX sequence was cloned into the GV303/EGFP lentiviral vector with two StrepⅡ tags and a single Flag tag at its C-terminal. Double restriction enzyme digestion was constructed correctly. Then MDA-MB-231 cells were transfected by the recombinant lentivirus vector. The cell line stably expression of fusion protein preliminary detected by real-time fluorescence quantitative PCR and Western Blot. The interaction between LOX and XRCC5 was detected by co-immunoprecipitation. In addition, we analyzed the distribution and interactions of XRCC5 with I,OX in MDA-MB-231 cells by indirect cell immunofluorescence assay. Results：The LOX-SF recombination protein expressed stably in MDA-MB- 231 cells. XRCC5 was detected in the protein complex from theimmunoprecipitationbyusinganti-LOX antibody. LOX also was detected in the immunoprecipitations by using anti-XRCC5 antibody. The co-localization of LOX and XRCC5 were positioned in nucleus by indirect cell immunofluorescence assay. Conclusion：The interaction between LOX and XRCC5 in human breast cancer cells was confirmed and the co-localizations were positioned in nucleus.