CAJ | 학술논문

目的探讨GLI3基因与单纯性马蹄内翻足(idiopathic congenital talipes equinovarus,ICTEV)的相关性.方法应用变性梯度凝胶电泳技术检测GLI3基因编码区的突变.用逆转录-PCR方法研究GLI3基因在1CTEV患者下肢的表达情况.构建ICTEV大鼠模型,应用实时定量PCR、免疫组织化学染色和蛋白质免疫印迹(Western blotting)方法研究Gli3基因在ICTEV模型鼠下肢肌肉组织中的表达.结果在84例ICTEV患者外周静脉血中未发现GLI3基因第1～8外显子以及第13外显子存在突变.在ICTEV患者及正常人下肢拇长屈肌中均未检测到GLI3基因的表达.不论在mRNA水平还是在蛋白质水平,Gli3基因在ICTEV模型胎鼠下肢组织中的表达均明显高于正常对照胎鼠.结论 GLI3基因的编码区突变可能不是ICTEV发病的主要原因,但GLI3基因的表达异常与马蹄内翻足的发病可能有关.
목적 탐토GLI3기인여단순성마제내번족(idiopathic congenital talipes equinovarus,ICTEV)적상관성.방법 응용변성제도응효전영기술검측GLI3기인편마구적돌변.용역전록-PCR방법연구GLI3기인재1CTEV환자하지적표체정황.구건ICTEV대서모형,응용실시정량PCR、면역조직화학염색화단백질면역인적(Western blotting)방법연구Gli3기인재ICTEV모형서하지기육조직중적표체.결과 재84례ICTEV환자외주정맥혈중미발현GLI3기인제1～8외현자이급제13외현자존재돌변.재ICTEV환자급정상인하지무장굴기중균미검측도GLI3기인적표체.불론재mRNA수평환시재단백질수평,Gli3기인재ICTEV모형태서하지조직중적표체균명현고우정상대조태서.결론 GLI3기인적편마구돌변가능불시ICTEV발병적주요원인,단GLI3기인적표체이상여마제내번족적발병가능유관.
[Objective]To investigate the relationship between GLI3 gene and pathogenesis of idiopathic congenital talipes equinovarus (ICTEV).[Methods] Potential mutations in the coding region of GLI3 were detected among 84 patients with ICTEV by denaturing gradient electrophoresis.Expression of GLI3 in the ICTEV patients' disease tissues was assessed by reverse transcription-PCR.Following generation of rat model for ICTEV,mRNA and protein levels of GLI3 were evaluated by real-time PCR and immunohistochemistry and Western blotting.[Results] No mutation was found in exons 1-8 and 13 of GLI3 gene among the 84 ICTEV patients.No expression of GLI3 gene was detected in the flexor hallucis longus of ICTEV patients or normal controls.Expression of Gli3,in terms of both mRNA and protein,was stronger in the hindlimb of ICTEV rat embryos compared with normal controls.[Conclusion] Mutation in the coding region of GLI3 may not be responsible for the occurrence of ICTEV.However,there may still be connection between abnormal expression of the gene and pathogenesis of ICTEV.