胰腺癌组织中BRAF表达的初步研究-东南大学学报医学版

胰腺癌组织中BRAF表达的初步研究

单位：东南大学附属中大医院,普外科,江苏,南京,210009

分类号：R735.9, R-33

出版年·卷·期（页码）：2006·25·第五期（333-336）

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 chip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Western blot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±0.00109,上调2.56倍;Western blot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 Western blot.Results The quantity of BRAF mRNA in normal pancreas and pancreatic adenocarcinoma tissues were(0.001 016 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 be(evaluated) in the future.

参考文献：

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