摘要
目的研究DNA修复酶基因NEIL1编码区发生的A-I RNA编辑是否在正常和癌细胞或组织中存在差异,并分析编辑事件的潜在影响,初步探究NEIL1的编辑事件与癌症的关联。方法通过RT-PCR和PCR分别扩增NEIL1编码区的目的片段,利用基于DNA和RNA扩增产物的测序峰图识别A-I RNA编辑并估算其编辑效率,采用T检验分析编辑效率在正常和癌细胞间差异是否具有统计学意义。结果在正常乳腺和乳腺癌细胞系,以及正常人和白血病患者白细胞中均检测到NEIL1上chr15-+-75646086和chr15-+-75646087两邻近位点发生A-I RNA编辑,前者导致NEIL1蛋白第242位的赖氨酸(K)转变为精氨酸(R)。与相应的正常细胞相比,在白血病患者白细胞中chr15-+-75646086位点的编辑水平显著下降,而在乳腺癌细胞系中chr15-+-75646087位点的编辑水平下降。结论 NEIL1编码区发生的A-I RNA编辑事件在白血病患者白细胞和乳腺癌细胞系中明显下降,提示编辑事件可能与癌症的发生发展相关。
Objective To uncover the role of RNA editing in cancer by detecting the editing level difference of two A-I RNA editing sites in NEIL1 gene between normal and tumor cells and analyzing the potential function of these implication editing events.Methods The identification of the A-I RNA editing event and a comparison of the editing level were carried out by comparing the cDNA sequences with their corresponding genomic templates based on PCR,RT-PCR and Sanger sequencing.Finally,T-test was used to detect difference of statistical significance between normal and cancer cells.Results In breast cancer,leukemia and corresponding normal cells,two A-I RNA editing sites mapped to chr15-＋-75646086 and chr15-＋-75646087 in NEIL1 were detected.The RNA editing site in chr15-＋-75646086 changed the lysine residue at position 242 of NEIL1 protein into arginine residue.Compared with the corresponding normal controls,the editing level of site chr15-＋-75646086 was significantly decreased in leukemia and the editing level of site chr15-＋-75646087 was decreased in breast cancer cell lines.Conclusion Down-regulation of RNA editing of NEIL1 in leukemia and breast cancer may be associated with the occurrence and progression of cancer.
出处
《军事医学》
CAS
CSCD
北大核心
2014年第6期458-462,共5页
Military Medical Sciences
基金
国家自然科学基金资助项目(31100940)
国家"重大新药创制"科技重大专项资助项目(2012ZX09J12106-03B)
