【摘要】：目的探討同源異形盒基因轉(zhuǎn)錄起始點(diǎn)(TSS)上游區(qū)域甲基化水平與神經(jīng)管缺陷(NTDs)的關(guān)系。方法采用兩階段病例對(duì)照研究設(shè)計(jì)。第一階段選擇NTDs病例10例和對(duì)照8例作為研究對(duì)象,運(yùn)用全基因組甲基化芯片檢測(cè)其腦或脊髓DNA全基因組甲基化水平。針對(duì)HOXA5基因差異甲基化區(qū)域,在第二階段擴(kuò)大樣本量(52例病例和23例對(duì)照)進(jìn)行驗(yàn)證。提取腦或脊髓組織DNA,使用Mass ARRAY系統(tǒng)的基質(zhì)輔助激光解吸電離飛行時(shí)間質(zhì)譜技術(shù)對(duì)HOXA5基因差異性甲基化區(qū)域進(jìn)行驗(yàn)證。結(jié)果第一階段在全基因組甲基化芯片結(jié)果中篩選出HOXA5基因TSS上游區(qū)27個(gè)CpG位點(diǎn)在病例組甲基化水平高于對(duì)照組(P0.05)。第二階段利用Mass ARRAY系統(tǒng)針對(duì)上述發(fā)現(xiàn)的差異區(qū)域進(jìn)行檢測(cè),成功檢測(cè)到10個(gè)可用于分析CpG位點(diǎn)。NTDs病例組及其亞型脊柱裂組和無腦畸形組中甲基化水平高于對(duì)照組的CpG位點(diǎn)數(shù)各有7、6和2個(gè)(P0.005)。HOXA5基因TSS上游區(qū)域病例組平均甲基化水平高于對(duì)照組[NTDs病例組:(31.3±13.9)%,對(duì)照組:(21.4±9.7)%],調(diào)整胎兒性別和孕周后OR值為1.09(1.03~1.16)。結(jié)論 HOXA5基因TSS上游區(qū)域高甲基化與胎兒神經(jīng)管缺陷風(fēng)險(xiǎn)增加存在關(guān)聯(lián)性。
[Abstract]:Objective to investigate the relationship between the level of methylation in the upstream region of (TSS) and neural tube defect (NTDs) at the transcriptional initiation point of homeobox gene. Methods A two-stage case-control study was designed. In the first stage, 10 cases of NTDs and 8 cases of control were selected as the subjects, and the whole genome methylated level of DNA in brain or spinal cord was detected by whole genome methylating chip. According to the differentially methylated region of HOXA5 gene, the sample size (52 cases and 23 controls) was expanded in the second stage. DNA, was extracted from brain or spinal cord tissue by matrix-assisted laser desorption ionization time-of-flight mass spectrometry with Mass ARRAY system to verify the differentially methylated region of HOXA5 gene. Results in the first stage, 27 CpG loci in the upstream region of HOXA5 gene TSS were screened out from the whole genome methylated chip results, and the methylation level in the case group was higher than that in the control group (P 0.05). In the second stage, the Mass ARRAY system is used to detect the difference areas found above. Ten CpG loci were successfully detected for the analysis of CpG loci. The number of CpG loci in NTDs case group and its subtypes of spina bifida group and anencephalic malformation group were 7, 6 and 2, respectively (P0.005). The upstream region of TSS of NTDs gene was 7, 6 and 2 (P0.005), respectively, which could be used to analyze NTDs and its subtypes of spina bifida and cerebral malformation. The average methylated level in the case group was higher than that in the control group [NTDs case group: (31.3 鹵13.9)%, In the control group: (21.4 鹵9.7)%, the adjusted fetal sex and OR value after gestational age were 1.09 (1.03 鈮，

本文編號(hào)：2474880