胰腺癌组织中BRAF表达的初步研究  

Study of the BRAF gene expression in pancreatic adenocarcinoma

作　　者：程张军[1] 石欣[1] 卫文俊[1] 汤永辉[1] 高乃荣[1] 

机构地区：[1]东南大学附属中大医院普外科,江苏南京210009

出　　处：《东南大学学报（医学版）》2006年第5期333-336,共4页Journal of Southeast University(Medical Science Edition)

基　　金：国家自然科学基金资助项目(30500491)

摘　　要：目的:了解BRAF基因在正常胰腺和胰腺癌组织中的表达水平。方法:6例胰腺癌标本取自胰腺癌手术患者,3例正常胰腺组织取自外伤性胰腺损伤需切除部分胰腺者。用Trizol法抽提每份组织的总RNA,分别进行紫外分光光度计测定、琼脂糖凝胶电泳及Lab on ch ip检测,对总RNA进行质控;应用实时定量PCR技术检测正常胰腺和胰腺癌组织BRAF的mRNA水平;应用W estern b lot检测BRAF的蛋白表达水平。结果:BRAF mRNA水平在正常胰腺组织中为0.001 016 7±0.000 25,胰腺癌组织中为0.002 606 2±0.001 09,上调2.56倍;W estern b lot显示,正常胰腺组织BRAF蛋白表达为6.70±2.50,胰腺癌组织为21.23±5.60,上调3.17倍。结论:胰腺癌组织中BRAF mRNA和蛋白表达水平均明显上调,其可能的调控机制及生物学效应还有待进一步研究。Objective To investigate the expression of the BRAF gene in human normal pancreas and pancreatic adenocarcinoma tissues. Methods Pancreatic adenocarcinoma tissue was obtained from 3 male and 3 female patients undergoing pancreas resection. Normal pancreatic tissue was obtained from 2 male and 1 female patients whose pancreas was injured by trauma. Total RNA were extracted from each sample with TRIzol method. To assure the quality of obtained total RNA, three different examinations were performed which included spectrophotometer, agarose gel electrophoresis and Lab on chip examination. The level of BRAF mRNA in normal pancreas and pancreatic adenocarcinoma tissues were detected by real-time fluorescence quantitative PCR. The protein level of BRAF was investigated using Westem blot. Results The quantity of BRAF mRNA in normal pancreas and pancreatic adenocarcinoma tissues were O. 001 016 7 ±0. 000 25 and 0. 002 606 2 ±0. 001 09, respectively. In comparison with normal pancreas, BRAF mRNA in pancreatic adenocarcinoma was overexpressed as 2.56 folds; Western blot showed that BRAF protein was 6.70± 2.50 in normal pancreas and 21.23 ± 5.60 in pancreatic adenocarcinoma, it was also overexpressed significantly in pancreatic cancer tissue by 3.17 folds. Conclusion BRAF mRNA and protein are significantly overexpressed in pancreatic adenocarcinoma. However, the regulatory mechanism of its overexpression and its real role in pancreatic adenocarcinoma has to he evaluated in the future.

关 键 词：胰腺癌 BRAF 实时定量聚合酶链反应 WESTERN BLOT 

分 类 号：R735.9[医药卫生—肿瘤] R-33[医药卫生—临床医学]