脆性X智力低下基因的体外封闭  

Silencing of FMR1 gene in vitro.

作　　者：张传仓[1] 陈姚[2] 姚英民[1] 

机构地区：[1]第一军医大学南方医院儿科,510515 [2]第一军医大学学员一旅

出　　处：《中国优生与遗传杂志》2003年第1期30-31,共2页Chinese Journal of Birth Health & Heredity

摘　　要：目的　初步建立一种FMR1基因的体外封闭方法 ,并初步探讨将其用于脆性X综合征研究的可能性 ;方法　利用NO可以诱导FMR1基因启动子区CpG岛甲基化的原理 ,将人类新鲜的外周血单个核细胞在含有 5 0 0 μmol/L硝普钠及2 0 %新生牛血清的RPMI 16 4 0培养基中培养 ,诱导FMR1基因启动子区CpG岛甲基化 ,从而使FMR1基因封闭 ,通过逆转录PCR ,检测FMR1基因的表达 ;结果　培养后第 12h、2 4h、4 8h均未检测到FMR1基因的表达 ,第 72h以后 ,重新表达 ,若于培养满 4 8h及时更换培养液 ,则培养第 72h亦未见表达 ;结论　本方法可在体外有效地封闭人类外周血单个核细胞的FMR1基因 ,模拟了脆性X综合征患者细胞的基因特点 。Objective: To establish a sort of in vitro gene silencing method of FMR1 gene,and to discuss the probability of its application in studies.Methods: Nitric oxide(NO) can induce methylation of the CpG island in the promoter region of FMR1 gene.Human fresh PBMC was cultured in FMRI?1640 containing 20% heat inactivated newborn bovine serum and 500μmol/L sodium nitroprusside(SNP,a kind of NO donor).RT?PCR was used to detect the expression of FMR1 gene.Results: No expression was found in the PBMC cultured with SNP at 12th,24th and 48th hour.But a positive band was found at 72th hour.If we changed the culture medium(containing SNP) at 48th hour,no positive band was found at 72th hour.Conclusion: FMR1 gene can be effectively silenced by this method in PBMC.That mimics the gene situation of the Fra(X) patients.So this method might be used in the studies of Fra(X) syndrome.

关 键 词：脆性X综合征 基因封闭 智力低下 

分 类 号：R596.1[医药卫生—内科学]