CAJ | 학술논문

目的系统分析22q11.2区基因在胚肾中的表达规律,并在患者中筛查候选基因的潜在突变.方法用逆转录-聚合酶链反应测定33个人类基因的同源体在不同胚龄以及成年小鼠肾脏中的表达水平,选取其中具有特异性表达者,在44例泌尿生殖系统畸形以及220名正常对照中进行突变筛查或排除,并对疑似突变者进行测序验证.为了减少误差,基因表达检测以及测序验证均重复两次.结果 K-means聚类分析提示,9个基因在胚肾发育全程均无表达;18个基因自发育初期或稍晚开始持续表达、强度仅呈微弱波动;6个基因在胚肾发育中呈短暂的一过性表达.结合文献研究,从后者中选择SNAP29基因进行全编码区突变筛查.聚合酶链反应-单链构象多态性及测序分析提示,在上述病例中,1例(隐睾)具有SNAP29基因第2外显子内的错义突变(GAG→AAG),2例(分别为隐睾和尿道下裂)在第3外显子中存在错义突变(AGC→GGC).对照组则未见相同突变.结论初步阐明了22q11.2区基因在小鼠胚肾发育期的表达规律,并证实多名泌尿生殖系统畸形患者具有候选基因SNAP29的错义突变,提示后者可能在泌尿生殖系统的发育中扮演关键角色.
목적 계통분석22q11.2구기인재배신중적표체규률,병재환자중사사후선기인적잠재돌변.방법 용역전록-취합매련반응측정33개인류기인적동원체재불동배령이급성년소서신장중적표체수평,선취기중구유특이성표체자,재44례비뇨생식계통기형이급220명정상대조중진행돌변사사혹배제,병대의사돌변자진행측서험증.위료감소오차,기인표체검측이급측서험증균중복량차.결과 K-means취류분석제시,9개기인재배신발육전정균무표체;18개기인자발육초기혹초만개시지속표체、강도부정미약파동;6개기인재배신발육중정단잠적일과성표체.결합문헌연구,종후자중선택SNAP29기인진행전편마구돌변사사.취합매련반응-단련구상다태성급측서분석제시,재상술병례중,1례(은고)구유SNAP29기인제2외현자내적착의돌변(GAG→AAG),2례(분별위은고화뇨도하렬)재제3외현자중존재착의돌변(AGC→GGC).대조조칙미견상동돌변.결론 초보천명료22q11.2구기인재소서배신발육기적표체규률,병증실다명비뇨생식계통기형환자구유후선기인SNAP29적착의돌변,제시후자가능재비뇨생식계통적발육중분연관건각색.
Objective To analyze the expression of genes from chromosomal region 22q11.2 and assess the association between mutation(s) of particular gene(s) from this region and malformations of the urinary system. Methods Expression of rat homologs of 33 genes from above region was determined in kidney tissues derived from rats of different fetal development ages (E13, E15, E19) and adulthood with reverse transcriptase-PCR. Potential mutation (s) in candidate gene SNAP29, whose expression pattern appeared to be unique, was screened in 44 patients and 220 normal controls with PCR-single strand conformation polymorphism(SSCP). Suspected positive regions were sequenced to verify the mutations. Results Nine genes showed no expression throughout the whole development process; 18 genes with various expression levels showed continuous expression from the beginning of development; 6 genes only expressed for a short time, among which SNAP29 was selected for mutation screening. Upon sequencing, three mutations were identified from the 44 patients, including a G to A transition (GAG to AAG) in exon 2, and two A to G transitions (AGC to GGC) in exon 3. Conclusion Through systematic analysis of the expression of genes from chromosomal region 22q11.2, the SNAP29 gene was found to have a potential role in the development of genitourinary system. Two missense mutations were identified in three patients. These included one in exon 2 (featuring cryptorchidism), and the other in exon 3 (featuring eryptorchidism and hypospadia). Neither of the mutations was found in the normal controls. The results suggested that mutation(s) of gene(s) from chromosomal region 22q11.2 may play an important role in the genesis of genitourinary malformations.