摘要
目的 通过对经逆行胰胆管造影(ERCP) 所获胰液的Kras 基因突变检测,以探索胰腺癌诊断的新方法。方法 应用聚合酶链反应(PCR)限制性片段长度多态性的方法检测胰腺良、恶性疾病组织物、胰液上清和细胞的Kras 突变。结果 胰腺癌和胰腺良性疾病标本分别有80% 和33 %的Kras 突变率。胰液上清检测时,PCR有18% 未获得成功,胰腺癌胰液Kras 突变率为71% (15/21),而胰腺良性疾病无一例有Kras 的突变。用胰液细胞有4 % PCR 未能获得成功,胰腺癌胰液细胞Kras的突变率为65% (11/17) ,良性疾病中有1 例(1/7) 检测到Kras 突变。综合上清与细胞的结果,在78 %(21/27) 的胰腺癌患者的胰液中检测到Kras 突变,明显高于胰腺良性疾病患者的8% (1/13)(P<0-001)。Kras 的突变率与肿瘤的部位及大小无统计学差异。结论 应用胰液检测Kras 突变有助于胰腺癌的诊断。
Objective Pancreatic neoplasm, which is considered as one of the most malignant tumors in humans, is now increasing steadily in frequency. The 5 year survival rate is less than 5%. In order to detect it in early stage, we tried to analyze K ras mutation in pancreatic juice collected during ERCP after injection of secretin. Mutation of coden 12 in exon 1 of K ras gene was reported as high as 90% in pancreatic cancer, but much less in benign pancreatic disease. Methods 47 patients at Peking Union Medical College Hospital between 1994 and 1998 were enrolled. Pancreatic juice collected during ERCP after injection of 100u secretin , was briefly centrifuged and the sediment was resuspended with PBS. Pancreatic juice supernatant and pancreatic cells were stored at -80℃ separately. DNA of the cells and supernatant of the juice were extracted. K ras point mutation was investigated in the DNA of these supernatant and cells by means of two stage polymerase chain reaction/restriction fragment length polymorphism (PCR/RFLP). Results When the supernatant of the pancreatic juice was used, the PCR successful rate was 82% , K ras mutation rate of the pancreatic carcinomas was 71% (15/21) compared with 0% (0/7) in benign pancreatic disease (3 chronic pancreatitis, 3 insulinoma and 1 pancreatic cyst). When the centrifuged pancreatic cells were used, the successful rate increased to 96%, sixty five percent (11/17) of pancreatic carcinomas had K ras mutation; mutation was also detected in 1/7 of the cases with benign pancreatic diseases (an insulinoma). In total, 78% (21/27) of the cases with pancreatic carcinomas has mutant alleles compared with 8% (1/13) of the benign pancreatic diseases( P <0.001) when pancreatic juice was used to detect K ras gene point mutation. In terms of the location of the pancreatic cancer, K ras mutation rate was not statistically different between the head (80%) and the body + tail(75%). The mutation rate did not vary with the size of the tumor. Conclusion Detection of coden 12 of K ras mutation in pancreatic juice can be used in differentiating pancreatic carcinomas from benign pancreatic diseases. It has high sensitivity and specificity.
出处
《中华内科杂志》
CAS
CSCD
北大核心
1999年第10期673-676,共4页
Chinese Journal of Internal Medicine
关键词
胰腺肿瘤
胰液
RAS基因
Pancreatic neoplasms Pancreatic juice Gene, ras