Gong X, Chen Y, Chen Y, Lu X
Department of Gastroenterology, Peking Union Medical College Hospital, Beijing 100730.
Zhonghua Nei Ke Za Zhi. 1999 Oct;38(10):673-6.
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.
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 degrees C 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).
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.
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.
胰腺癌被认为是人类最恶性的肿瘤之一,其发病率目前正稳步上升。5年生存率低于5%。为了早期检测胰腺癌,我们尝试分析在注射促胰液素后经内镜逆行胰胆管造影(ERCP)收集的胰液中的K-ras突变。K-ras基因第1外显子12密码子的突变在胰腺癌中报道高达90%,而在良性胰腺疾病中则少得多。
选取1994年至1998年在北京协和医院就诊的47例患者。在注射100单位促胰液素后经ERCP收集胰液,短暂离心,沉淀物用磷酸盐缓冲液(PBS)重悬。胰液上清液和胰腺细胞分别保存在-80℃。提取细胞和胰液上清液的DNA。通过两阶段聚合酶链反应/限制性片段长度多态性(PCR/RFLP)方法研究这些上清液和细胞DNA中的K-ras点突变。
当使用胰液上清液时,PCR成功率为82%,胰腺癌的K-ras突变率为71%(15/21),而良性胰腺疾病(3例慢性胰腺炎、3例胰岛素瘤和1例胰腺囊肿)的K-ras突变率为0%(0/7)。当使用离心后的胰腺细胞时,成功率提高到96%,65%(11/17)的胰腺癌有K-ras突变;在1/7的良性胰腺疾病病例(1例胰岛素瘤)中也检测到突变。当用胰液检测K-ras基因点突变时,胰腺癌病例中有78%(21/27)有突变等位基因,而良性胰腺疾病为8%(1/13)(P<0.001)。就胰腺癌的位置而言,胰头癌(80%)和胰体尾癌(75%)的K-ras突变率无统计学差异。突变率不随肿瘤大小而变化。
检测胰液中K-ras第12密码子突变可用于鉴别胰腺癌与良性胰腺疾病。其具有高敏感性和特异性。