CAJ | 학술논문

目的探讨双向荧光差异凝胶电泳筛选胆管癌胆汁差异表达蛋白的可行性.方法实验组与对照组胆汁各6例,分别取自胆管癌及胆总管结石病例.实验与对照组每例各取100μg蛋白质,分别标记400 pmol的Cy3与Cy5,另每例各取50 μg混合后制备成内标样品并标记2.4 nmol的Cy2.标记后的实验组、对照组及内参组样品各取100 μg等量混合后在同一凝胶内进行双向荧光差异凝胶电泳,通过质谱分析,筛选并鉴定有统计学意义的差异表达蛋白,探讨其生物学意义.结果实验组与对照组共筛选出55个差异表达蛋白,其表达量两组间相差1.5倍以上,t检验有统计学意义(P＜0.05).对其中15个差异表达蛋白进行质谱鉴定,共鉴定出8种差异表达蛋白.结论双向荧光差异凝胶电泳胆汁中筛选胆管癌胆汁差异表达蛋白是可行的,可用于胆管癌早期诊断标志物的筛选.
목적 탐토쌍향형광차이응효전영사선담관암담즙차이표체단백적가행성.방법 실험조여대조조담즙각6례,분별취자담관암급담총관결석병례.실험여대조조매례각취100μg단백질,분별표기400 pmol적Cy3여Cy5,령매례각취50 μg혼합후제비성내표양품병표기2.4 nmol적Cy2.표기후적실험조、대조조급내삼조양품각취100 μg등량혼합후재동일응효내진행쌍향형광차이응효전영,통과질보분석,사선병감정유통계학의의적차이표체단백,탐토기생물학의의.결과 실험조여대조조공사선출55개차이표체단백,기표체량량조간상차1.5배이상,t검험유통계학의의(P＜0.05).대기중15개차이표체단백진행질보감정,공감정출8충차이표체단백.결론 쌍향형광차이응효전영담즙중사선담관암담즙차이표체단백시가행적,가용우담관암조기진단표지물적사선.
Objective To determine the probability of identification of differential expression of biliary proteins induced by cholangiocarcinoma using 2D-DIGE. Methods Bile was obtained from 12patients with obstructive jaundice (including 6 cases of cholangiocarcinoma and 6 of cholelithiasis).Each sample was labeled with three different CyDyes (y3,Cy5,Cy2) including one internal standard,pooled from all the samples, and separated with 2-D DIGE in triplicate experiments. MALDI-TOF-MS and bioinformatics were adopted to identify and elucidate the significance of differentially expressed proteins in bile induced by cholangiocarcinoma. Results 55 matched protein spots differences in abundance were detected with statistical variance of two groups(Average Volum Ratio ≥1.5, t-test, P＜0. 05). Among these proteins, 13 PMF were obtained by MALDI-TOF-MS analysis. Eight proteins were identified by searching a protein database. Conclusion The differentially displayed proteomes between the pathological bile obtained from benign and malignant obstructive jaundice indicates the potential application of 2D-DIGE to identify the biomarker of cholangiocarcinoma.