Niedergethmann M, Rexin M, Knob S, Hartel M, Sturm J W, Richter A, Post S
Chirurgische Klinik, Universitätsklinikum Mannheim, Fakultät für klinische Medizin Mannheim der Universität Heidelberg, Germany.
Zentralbl Chir. 2001 Nov;126(11):917-21. doi: 10.1055/s-2001-19144.
Despite apparently curative resection adenocarcinomas of the pancreas early recur. Thus, the pathological examination should be enriched by sensitive methods to detect minimal residual disease (MRD). Mutant K-ras is the most promising genetic alteration in ductal adenocarcinoma and may serve to detect malignant cells by polymerase chain reaction (PCR) based techniques. Therefore, we set out to detect K-ras mutations by PCR for evaluation of MRD in patients after curative resection of pancreatic adenocarcinoma.
Tumor tissue and corresponding paraaortic lymph nodes were obtained from 51 patients, who underwent surgery for pancreatic head tumors. The paraaortic lymph nodes were staged as tumor-free by routine histopathology in all cases diagnosed for ductal adenocarcinoma (study group, n = 40) or other tumors (control group, n = 11). Therefore, DNA of both primary tumors and lymph nodes was extracted and analysed by a PCR-based assay with respect to mutated K-ras. As a positive control the human pancreatic cancer cell line PaTu-8902 was used.
K-ras mutations were detected in 73 % (29/40) of primary tumors of ductal adenocarcinomas and in 17 % (5/29) in the corresponding paraaortic lymph nodes, which were diagnosed as tumor-free by routine pathology. The identical type of point mutation was found in primary tumors and corresponding lymph nodes by use of sequence specific primers. In the control group no K-ras mutation was detected.
Tumor cell DNA can be detected sensitively in tumor- and lymph node specimen with the described method. Routinely assessed, this method is able to detect MRD and could enrich the pathological examination, in order to determine prognostic relevant subgroups of patients.
尽管胰腺癌看似根治性切除后仍早期复发。因此,病理检查应采用敏感方法以检测微小残留病(MRD)。突变型K-ras是导管腺癌中最有前景的基因改变,可通过基于聚合酶链反应(PCR)的技术用于检测恶性细胞。因此,我们着手通过PCR检测K-ras突变,以评估胰腺癌根治性切除术后患者的MRD。
从51例行胰头肿瘤手术的患者获取肿瘤组织及相应的腹主动脉旁淋巴结。在所有诊断为导管腺癌的病例(研究组,n = 40)或其他肿瘤(对照组,n = 11)中,腹主动脉旁淋巴结经常规组织病理学检查均判定无肿瘤。因此,提取原发性肿瘤和淋巴结的DNA,并通过基于PCR的检测方法分析突变型K-ras。使用人胰腺癌细胞系PaTu-8902作为阳性对照。
在导管腺癌原发性肿瘤中,73%(29/40)检测到K-ras突变,在相应的腹主动脉旁淋巴结中,17%(5/29)检测到K-ras突变,这些淋巴结经常规病理学检查诊断为无肿瘤。通过使用序列特异性引物,在原发性肿瘤和相应淋巴结中发现相同类型的点突变。在对照组中未检测到K-ras突变。
使用所述方法可在肿瘤和淋巴结标本中灵敏地检测到肿瘤细胞DNA。经常规评估,该方法能够检测MRD,并可丰富病理检查,以确定患者的预后相关亚组。