十二指肠液K-ras癌基因检测在胆管癌早期诊断中应用价值.docVIP

十二指肠液K-ras癌基因检测在胆管癌早期诊断中应用价值 　【摘要】 目的 探讨十二指肠液标本中K-ras癌基因检测对胆管癌早期诊断的价值。方法 应用DNAzolR 试剂提取DNA，通过聚合酶链反应-限制性片段长度多态性（polymerase chain reaction restriction fragment length polymorplism，PCR-RFLP）法分析，检测30例胆管癌患者和10例良性胆道疾病患者胆汁及十二指肠液标本中上清液及沉渣的K-ras基因突变情况。结果 胆管癌组胆汁标本的上清液中K-ras基因突变检测的阳性率为56.7%；相应的十二指肠液标本阳性率为43%。良性胆道疾病组仅胆汁标本检出K-ras基因突变1例，良恶性疾病组比较差异有统计学意义﹙Plt;0.005），胆管癌K-ras基因突变的检出率在上清液组中明显高于沉渣组，两者比较差异有统计学意义﹙Plt;0.005），对于胆管刷检阴性的胆管癌患者，检测胆汁中K-ras基因突变的阳性率仍可达39.3%。结论 应用DNAzolR 试剂能有效提取到胆汁及十二指肠液标本中的DNA，十二指肠液K-ras基因检测可能有助于胆管癌的早期诊断。 【关键词】 胆管肿瘤 K-ras基因 十二指肠液 Abstact Objecive To explore the applied value of detection of K-ras gene mutations in the duodenal juice of patients with biliary tract carcinoma (BTCa) in early diagnosis. Methods DNA was extracted using DNAzolR reagent. The situation of K-ras mutation in the supernant and sediments in the sample of bile and duodenal in 30 cases with BTCa and 10 cases with benign choledocholithiasis were detected with PCR-RFLP. Results The positive rates of assay with K-ras gene mutations in bile supernatant of patients with BTCa were 56.7%. Accordingly， the positive rates of those in duodenal juice were 43%. Only one case K-ras gene mutation was detected in benign choledochlithiasis. Statistic analysis showed there was significant difference in those gene mutation between BTCa group and benign group(Plt;0.005). Positive rate of K-ras gene mutations was higher in the supernatant than that in the sediments (Plt;0.005). For the negative of bile duct brushing in BTCa, positive rates of assay with K-ras gene mutation could still reach 39.3%. Conclusions DNA can be extracted from bile and duodenal juice with DNAzolR reagent. Assay with K-ras gene mutations in duodenal juice may be useful for early diagnosis of biliary tract carcinoma. Key words biliary tract neoplasms； K-ras gene； duodenal juice 胆管癌系高度恶性肿瘤，以往认为其发病率低，近年来随着现代医学诊疗技术的发展，胆管癌的发病率也不断上升，同时对此病的诊治也不断取得进展，但由于胆管癌的解剖部位隐匿，早期诊断仍非常困难，一旦发现多为晚期，致使手术切除率低，预后差。发展新的筛选和诊断手段，是提高胆管癌早期诊断率和改善胆管癌预后的关键。 我们采用分子生物学技术