肺癌细胞中CD59 rs79077373遗传变异对其转录活性的影响

Functional analysis of CD59 rs79077373 in lung cancer cells

目的探讨肺癌细胞中CD59启动子区rs79077373遗传变异对其转录活性的影响。方法利用TRANSFAC预测可能影响CD59转录活性的启动子区遗传变异,然后构建含有不同等位基因的CD59启动子区报告基因载体,分别命名为pGL3-rs79077373C和pGL3-rs79077373T,将重组质粒与内参质粒pRL-SV40共转染于肺癌细胞A549、NCI-H23、NCI-H2030后,检测报告基因活性。结果生物信息学分析发现当CD59 rs790773737增加转录因子TBP的结合。荧光素酶报告基因结果显示,在肺癌细胞A549、NCI-H2030、NCI-H23细胞中pGL3-rs79077373T组的相对荧光素酶活性分别是pGL3-rs79077373C组的2.00、1.82和1.42倍(P <0.05)。结论 CD59启动子区rs79077373 C>T变异可增加CD59的启动子的活性。

Objective To investigated the effect of rs79077373 （C/T） mutation in promoter region of CD59 on CD59 transcription activity. Methods Bioinformatic analysis was performed to predict potential mutation in transcription factor binding site of the promoter region of CD59 that may affect transcription activity. Two different pGL3-CD59 promoter luciferase reporter constructs, CD59 rs790773737 T and pGL3-rs79077373C, were generated. Constructers were co-transfected with pRL-SV40 plasmid into A549, NCI-H2030 and NCI-H23 cells for manifestation of the transcription activity of CD59. Results Bioinformatic analysis unveiled that alteration of CD59 rs79077373 T allele to C allele may affect the transcription activity. The reporter gene expression suggested a 2.00-fold, 1.82-fold and 1.42-fold increase of transcription activity in A549, NCI-H2030 and NCI-H23 cells in CD59 rs79077373 T allele group compared with CD59 rs79077373C group （respectively, P 〈 0.05）.Conclusions CD59rs79077373 carrying the T allele enhances CD59 transcriptional activity.