CAJ | 학술논문

目的分析DIXDC1和AXIN2基因在先天性巨结肠症(hirschsprung disease,HSCR)中的表达,探讨其在HSCR中的意义.方法采用蛋白印迹(Western blot)、免疫组化和荧光实时定量PCR(quantitative real-time PCR,qRT-PCR)方法检测60例HSCR患儿正常段(有神经节段肠管)和狭窄段(无神经节段肠管)中DIXDC1和AXIN2的表达,并对其表达进行定量与比较分析.结果 DIXDC1和AXIN2在HSCR狭窄段肠管中蛋白相对表达量为27.16±2.04和29.63±4.72,高于正常段肠管中的13.27±3.15和14.99±2.82,差异有统计学意义(DIXDC 1 P＜0.041;AXIN2 P＜0.035).DIXDC1和AXIN2在HSCR狭窄段肠壁的肌间和黏膜下细胞胞质内呈强阳性反应,而在HSCR正常段肠壁的肌间和黏膜下细胞胞质内呈弱阳性或阴性.DIXDC1和AXIN2在HSCR狭窄段肠管中mRNA相对含量为22.34±1.23和25.77±2.06,高于正常段肠管中的12.53±1.47和13.98±1.62,差异有统计学意义(DIXDC1 P＜0.033;AXIN2 P＜0.029).结论 DIXDC1和AXIN2 mR-NA与蛋白在HSCR肠管组织中的异常表达,可能与HSCR的发生关系密切,并可能在先天性消化道畸形的肠道发育中起一定作用.
목적 분석DIXDC1화AXIN2기인재선천성거결장증(hirschsprung disease,HSCR)중적표체,탐토기재HSCR중적의의.방법 채용단백인적(Western blot)、면역조화화형광실시정량PCR(quantitative real-time PCR,qRT-PCR)방법검측60례HSCR환인정상단(유신경절단장관)화협착단(무신경절단장관)중DIXDC1화AXIN2적표체,병대기표체진행정량여비교분석.결과 DIXDC1화AXIN2재HSCR협착단장관중단백상대표체량위27.16±2.04화29.63±4.72,고우정상단장관중적13.27±3.15화14.99±2.82,차이유통계학의의(DIXDC 1 P＜0.041;AXIN2 P＜0.035).DIXDC1화AXIN2재HSCR협착단장벽적기간화점막하세포포질내정강양성반응,이재HSCR정상단장벽적기간화점막하세포포질내정약양성혹음성.DIXDC1화AXIN2재HSCR협착단장관중mRNA상대함량위22.34±1.23화25.77±2.06,고우정상단장관중적12.53±1.47화13.98±1.62,차이유통계학의의(DIXDC1 P＜0.033;AXIN2 P＜0.029).결론 DIXDC1화AXIN2 mR-NA여단백재HSCR장관조직중적이상표체,가능여HSCR적발생관계밀절,병가능재선천성소화도기형적장도발육중기일정작용.
Objective To study the expression of DIXDC1 and AXIN2 gene in tissue samples from the patients with Hirschsprung disease (HSCR) and validate their significane in HSCR.Methods Western blot,immunohistochemical staining and real-time quantitative PCR (qRT-PCR) methods were done in 60 cases of HSCR patients at the normal segment (ganglion section of bowel) and the stenotic segment (absence of ganglion section of bowel) for DIXDC1 and AXIN2 expression.Results The protein expression of DIXDC1 and AXIN2 in stenotic segment were 27.16 ± 2.04 and 29.63 ±4.72,as compared to normal segment (13.27 ± 3.15 and 14.99 ± 2.82).The difference was statistically significant (DIXDC 1 P＜0.041 and AXIN2 P＜0.035).In the myenteric ganglia,the submucosa,muscle and longitudinal muscle of ganglionic segment,immunohistochemical staining of DIXDC1 and AXIN2 were detected with low expression.High expression of these were found in aganglionic segment.The mRNA expression of DIXDC1 and AXIN2 were 22.34 ± 1.23 and 25.77 ± 2.06 respectively in stenotic segment,and they were higher than normal segment (12.53 ± 1.47 and 13.98 +1.62).The difference was also statistically significant (DIXDC 1 P＜0.043;AXIN2 P＜0.039).Conclusions DIXDC1 and AXIN2 genes and protein expression were higher in aganglionic segments in HSCR. This would suggest that DIXDC1 and AXIN2 were involved in the formation stages of HSCR,and may play a role in the pathogenesis congenital malformation of the alimentary tract.