中华胃肠外科杂志
中華胃腸外科雜誌
중화위장외과잡지
CHINESE JOURNAL OF GASTROINTESTINAL SURGERY
2010年
10期
758-761
,共4页
高红%张志波%姜中佳%王大佳%黄英%王维林
高紅%張誌波%薑中佳%王大佳%黃英%王維林
고홍%장지파%강중가%왕대가%황영%왕유림
先天性巨结肠%基因,WNT8b%基因%SHH%突变
先天性巨結腸%基因,WNT8b%基因%SHH%突變
선천성거결장%기인,WNT8b%기인%SHH%돌변
Hirschsprung disease%Gene,WNT8b%Gene,SHH%Mutation
目的 探讨WNT8b和SHH基因突变与中国儿童先天性巨结肠症(HSCR)发病的关系.方法 收集东北地区72例散发性HSCR患儿术前全血标本(病例组),同时选取72名性别、年龄相匹配的健康儿童外周血作为对照(对照组).提取上述外周血标本基因组DNA后,应用PCR方法对WNT8b基因第1外显子和SHH基因第1外显子进行基因突变的检测,将突变样品进行自动测序分析.采用荧光实时定量PCR(qRT-PCR)方法检测外周血WNT8b和SHH基因的mRNA水平.结果 病例组72例HSCR患儿的WNT8b基因测序结果显示,13例WNT8b基因编码区发生突变,其中杂合性缺失(A缺失)8例(11.1%),碱基置换突变5例(6.9%);SHH基因测序结果显示,11例SHH基因编码区发生突变,其中杂合性缺失(A缺失)7例(9.7%),碱基置换突变4例(5.6%);从而引发氨基酸的改变;而对照组均未发现上述突变.病例组与对照组外周血WNT8b和SHH mRNA表达水平分别为30.01±1.13、17.33±0.62和28.25±1.27、18.94±0.31,差异有统计学意义(均P＜0.05).结论 HSCR患儿外周血WNT8b和SHH基因存在突变和异常表达,这两种基因可能与东北地区中国儿童HSCR的发生有关.
目的 探討WNT8b和SHH基因突變與中國兒童先天性巨結腸癥(HSCR)髮病的關繫.方法 收集東北地區72例散髮性HSCR患兒術前全血標本(病例組),同時選取72名性彆、年齡相匹配的健康兒童外週血作為對照(對照組).提取上述外週血標本基因組DNA後,應用PCR方法對WNT8b基因第1外顯子和SHH基因第1外顯子進行基因突變的檢測,將突變樣品進行自動測序分析.採用熒光實時定量PCR(qRT-PCR)方法檢測外週血WNT8b和SHH基因的mRNA水平.結果 病例組72例HSCR患兒的WNT8b基因測序結果顯示,13例WNT8b基因編碼區髮生突變,其中雜閤性缺失(A缺失)8例(11.1%),堿基置換突變5例(6.9%);SHH基因測序結果顯示,11例SHH基因編碼區髮生突變,其中雜閤性缺失(A缺失)7例(9.7%),堿基置換突變4例(5.6%);從而引髮氨基痠的改變;而對照組均未髮現上述突變.病例組與對照組外週血WNT8b和SHH mRNA錶達水平分彆為30.01±1.13、17.33±0.62和28.25±1.27、18.94±0.31,差異有統計學意義(均P＜0.05).結論 HSCR患兒外週血WNT8b和SHH基因存在突變和異常錶達,這兩種基因可能與東北地區中國兒童HSCR的髮生有關.
목적 탐토WNT8b화SHH기인돌변여중국인동선천성거결장증(HSCR)발병적관계.방법 수집동북지구72례산발성HSCR환인술전전혈표본(병례조),동시선취72명성별、년령상필배적건강인동외주혈작위대조(대조조).제취상술외주혈표본기인조DNA후,응용PCR방법대WNT8b기인제1외현자화SHH기인제1외현자진행기인돌변적검측,장돌변양품진행자동측서분석.채용형광실시정량PCR(qRT-PCR)방법검측외주혈WNT8b화SHH기인적mRNA수평.결과 병례조72례HSCR환인적WNT8b기인측서결과현시,13례WNT8b기인편마구발생돌변,기중잡합성결실(A결실)8례(11.1%),감기치환돌변5례(6.9%);SHH기인측서결과현시,11례SHH기인편마구발생돌변,기중잡합성결실(A결실)7례(9.7%),감기치환돌변4례(5.6%);종이인발안기산적개변;이대조조균미발현상술돌변.병례조여대조조외주혈WNT8b화SHH mRNA표체수평분별위30.01±1.13、17.33±0.62화28.25±1.27、18.94±0.31,차이유통계학의의(균P＜0.05).결론 HSCR환인외주혈WNT8b화SHH기인존재돌변화이상표체,저량충기인가능여동북지구중국인동HSCR적발생유관.
Objective To investigate the relationship of WNT8b and SHH genes mutation and Hirschsprung disease(HSCR) in Chinese children. Methods Preoperative whole blood preparations in 72 children with sporadic HSCR from northeast China were collected(study group). Seventy-two healthy children were used as controls(matched for sex and age). Genomic DNA was obtained from peripheral blood. Exon 1 of WNT8b gene and the exon 1 of SHH gene were analyzed for gene mutation. The mutation products were automatically sequenced. The levels of WNT8b and SHH Mrna were detected by quantitative real-time PCR(Qrt-PCR) in blood samples. Results On sequencing, 13 out of 72 children with HSCR had WNT8b gene mutation in the coding area, including heterozygosity deletion in 8 cases (11.1%) and base replacement in 5(6.9%). Eleven children with HSCR had SHH gene mutation in the coding area including heterozygosity deletion in 7 cases(9.7%) and base replacement in 4(5.6%). No mutations in WNT8b and SHH genes were found in the control group. The WNT8b and SHH Mrna levels were different between the study group and the control group(30.01±1.13 vs. 17.33±0.62,and 28.25±1.27 vs. 18.94±0.31,P＜0.05). Conclusions WNT8b and SHH mutations and abnormal expressions are present in the peripheral blood of children with sporadic HSCR. These two genes may be related to the development of sporadic HSCR in children in the northeastern China.
