胰液端粒酶活性与K-ras检测对胰腺癌的诊断价值

Detection of telomerase activity and K-ras gene mutation in pancreatic juice and its diagnostic value in pancreatic cancer

目的 检测胰腺癌和慢性胰腺炎患者胰液中K ras基因突变和端粒酶活性 ,探讨端粒酶活性与K ras基因突变联合检测对胰腺癌患者诊断价值。方法 收集 2 4例胰腺癌和 14例慢性胰腺炎患者胰液 ,K ras基因突变采用聚合酶链反应 -限制性酶切片段长度多态性 ( polymerasechianreaction -restrictionfragmentlengthpolymorphism ,PCR -RFLP)检测 ,端粒酶活性检测采用端粒末端重复序列扩增技术 (telomericrepeatamplificationprotocol ,TRAP)。 结果 胰腺癌中K ras基因突变率为 87.5 % ( 2 1/2 4) ,慢性胰腺炎中K ras基因突变率仅为 2 1.3 % ( 3 /14 ) ,差异有显著性 (P<0 .0 1) ;在 2 4例胰腺癌中端粒酶阳性检出率为 83 .3 % ( 2 0 /2 4) ;而 14例慢性胰腺炎胰液中端粒酶阳性检出率为 2 8.6% ( 4 /2 4) ,差异同样具有显著性 (P <0 .0 1)。K ras基因突变与端粒酶活性检测诊断胰腺癌的特敏感性分别为 87.5 % ,83 .3 % ;两者联合检测诊断胰腺癌的敏感性为 85 .5 % ,特异性达到 85 .7%。结论 联合检测胰液中K ras基因与端粒酶活性可提高胰腺癌诊断的敏感性和特异性 ,对胰腺癌筛查。

Objective To detect K-ras gene mutation and telomerase activity in pancreatic juice of pancreatic cancer (PC) and chronic pancreatitis (CP),and evaluate their diagnostic value.Methods K-ras 12 codon mutation was detected by polymerase chian reaction-restriction fragment length polymorphism (PCR-RFLP) in 24 cases of pancreatic cancer and 14 cases of chronic pancreatitis, and telomerase activity was determined by SYBR GreenⅠ-staining telomeric repeat amplification protocol assay (TRAP).Results K-ras mutation was detected in 87.5%(21/24) of PC cases and 21.3%(3/14) of CP cases respectively with the difference being statistically significant(P<0.01).Telomerase activity was positive in 83.3%(20/24) of PC cases and 28.6%(4/14) of CP cases respectively with the difference being significant (P<0.01).The sensitivity of K-ras mution and telomerase activity assay was 87.5% and 83.3% respectively.The sensitivity and specificity of combined assay reached 85.5% and 85.7% respectively.Conclusion Combined detection of K-ras mutation and telomerase activity in pancreatic juice is helpful to screen and diagnose pancreatic carcinoma.