十二指肠液K-ras、P53和P16基因联合检测在胆管癌诊断中的价值

Clinical singnificance in diagnosis of biliary tract carcinoma by detection of K-ras、P53 and P16 gene mutations in duodenal juice

目的探讨十二指肠液标本中K-ras、P53和P16基因联合检测对胆管癌诊断的价值。方法应用PCR-SSCP-银染法和PCR-RFLP法检测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、P53及P16基因突变状况。结果胆管癌组胆汁标本的上清中K-ras、P53和P16基因突变及三基因联合检测的阳性率为56.7%、50%、33%和80%;相应的十二指肠液标本阳性率为43%、36%、20%和70%。良性胆道疾病组仅胆汁标本检出K-ras基因突变1例,良恶性疾病组比较有显著性差异(P<0.005),胆管癌3种基因突变的检出率在上清组中明显高于沉渣组(P<0.001),对于胆管刷检阴性的胆管癌患者,联合检测胆汁中3种基因突变的阳性率仍高达66.67%。结论十二指肠液K-ras、P53和P16基因联合检测有助于胆管癌的诊断。

Objective To investigate the role of K-ras, P53, and P16 gene of duodenal juice detection in diagnosis of biliary tract carcinoma (BTC). Methods The K-ras, P53 and P16 gene mutations in the bile and duodenal juice from 30 cases with BTC and 10 cases with benign choledocholithiasis were detected by PCR-SSCP-AgNO3 staining or PCR-RFLP. Results The positive rates of K-ras, P53 and P16 gene mutations and combination assay with them in bile supernatant of patients with BTC were 56%, 50%, 33% and 83.3%, respectively. The positive rates of them in duodenal juice were 43%, 36%, 20% and 70%, respectively. However, K-ras gene mutation was detected in only one case with benign choledochlithiasis. Statistic analysis show there was significant difference in those gene mutations between BTC group and benign group (P<0.001). Positive rates of K-ras, P53 and P16 gene mutations were higher in the supernatant than that in the sediment (P<0.001). For the negative of blie duct brushing in BTC, positive rate of combination assay with these gene mutations was still high (66.67%). Conclusions Combined assay of K-ras, P53 and P16 gene mutations in duodenal juice might be useful for diagnosis of BTC.