中文摘要：

目的构建重组X连锁凋亡抑制蛋白质（XIAP）基因3′非翻译区（3′UTR）荧光素酶报告载体,分析可能调控XIAP基因表达的微RNA（miRNA）。方法采用聚合酶链反应（PCR）从人cDNA中扩增XIAP-3′UTR序列,插入荧光素酶报告载体pGL3-Ctrl,获得重组载体pGL3-Ctrl/XIAP;采用Target Scan 6.2软件预测可能与XIAP-3′UTR结合的miRNA;将pGL3-Ctrl/XIAP重组质粒和miRNA共转染A549细胞,测定XIAP-3′UTR荧光素酶的活性。结果酶切及核酸测序证实,成功构建了XIAP-3′UTR序列的荧光素酶报告重组子;miRNA靶位点的预测显示,XIAP基因可能是miR-200b、miR-200c和miR-429的作用靶标;与miRNA mimic ctrl组比较,miR-200b、miR-200c和miR-429能明显降低pGL3-Ctrl/XIAP的荧光素酶活性,差异有统计学意义（P〈0.05）。结论成功构建了XIAP-3′UTR荧光素酶报告载体,且miR-200b、miR-200c和miR-429可显著降低其荧光素酶的活性。

英文摘要：

Objective To construct the recombinant X-linked inhibitor of apoptosis protein（XIAP） gene 3′untranslational region （3′UTR）-luciferase reporter vector ,and analyze the microRNA（miRNA） which possibly regulate the expression of XIAP gene . Methods Polymerase chain reaction （PCR） was employed to amplify X IA P-3′UTR sequences from human cDNA ,in which luciferase reporter vector pGL3-Ctrl was inserted ,and the recombinant vector pGL3-Ctrl/XIAP was gained .Target Scan 6 .2 soft-ware was adopted to predict the miRNA which possibly combined with the X IA P-3′UTR .pGL3-Ctrl/XIAP recombinant plasmids and the miRNA were co-transfected into A549 cells ,and the X IA P-3′UTR-luciferase activity was measured .Results Confirmed by digestion and DNA sequencing ,the X IA P-3′UTR-luciferase reporter recombinant was successfully constructed .Prediction of miRNA target sites indicated that X IA P gene may be the target of miR-200b ,miR-200c and miR-429 .Compared with miRNA mim-ic ctrl group ,miR-200b ,miR-200c and miR-429 significantly reduced the luciferase activity of pGL 3-Ctrl/XIAP with statistically significant difference（P〈0 .05） .Conclusion X IA P-3′UTR-luciferase reporter vector is successfully constructed .miR-200b ,miR-200c and miR-429 can obviously decrease the luciferase activity .