The molecular and clinicopathological study related to detection of K-ras point mutation in the blood of pancreatic cancer cases

胰腺癌患者血液中K-ras点突变检测相关的分子及临床病理学研究

基本信息

批准号：
08671480
负责人：
NAGAI Hideo
金额：
$ 1.47万
依托单位：
JICHI MEDICAL SCHOOL
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1996
资助国家：
日本
起止时间：
1996 至 1998
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08671480/
关键词：
pancreatic cancer peripheral blood K-ras point mutation PCR-RFLP magnetic cell sorting system allele specific PCR

项目摘要

Purpose : We have attempted to detect K-ras mutation from blood specimens in order to assist in the diagnosis of pancreatic cancer.Materials : The blood (perioperative peripheral blood and intraoperative portal blood) and tissues of the primary lesion in the pancreas were taken from 16 patients with pancreatic cancer (n=14) and benign diseases (n=2 ; chronic pancreatitis and insulinoma).Methods and Results :1.Detection of K-ras mutation in the primary lesionAfter microdissection we extracted DNA whose K-ras gene was amplified by PCR system. Its sequence was identified by digoxigenin dot blotting. The incidence of mutation at codon 12 was 85%.2.Detection of K-ras mutation from blood specimensBy the magnetic cell sorting system (MCSS) using anticytokeratine antibody and magnetic microspheres, we concentrated epithelial cells from the blood. Following amplification of the K-ras gene we enriched the mutated gene by the PCR-RFLP method. The sensitivity of PCR-RFLP without MCSS was only 10^< … More -4>, whereas that of combination of PCR-RFLP and MCSS reached 10^<-6> -10^<-7> With this combined method we could detect K-ras mutation in all samples of the portal blood and postoperative peripheral blood when the patients had K-ras mutation in the primary tumor. Furthermore, the mutation was detected in 25% even in the preope-rative peripheral blood. These cases rapidly deteriorated after surgery and died 3-6 months postoperatively.Discussion and ConclusionThe current study showed that tumor cells existed in the peripheral blood of pancreatic cancer patients at the rate of one tumor cell per 105 to 107 normal nucleated cells. We conjectured that the number of circulating cancer cells in the preoperative blood might be several thousand per body, and that the operative injury seemed to increase their number ten times. It was suggested that prognosis would be extremely poor if patients had tens of thousand of circulating cancer cells in the peripheral blood before surgey. We think that even the combination of PCR-RFLP and MCSS is not appropriate for early detection of pancreatic cancer. However, this system appears to be useful in excluding contraindicated operative cases from surgery. Less

目的：我们试图从胰腺癌诊断中从血液标本中检测。材料：血液（血液（术中外周血液和术中血液）和胰腺中原发性病变的组织和胰腺中原发性病变的组织来自16例胰腺患者癌症（n = 14）和良性探测。方法和结果：1。在初级病变后微解原下进行S突变的dna我们提取的DNA，其k-ras基因通过使用抗细菌抗体抗体的磁细胞系统（MCSS）扩增其K-RAS基因的k-ras基因被扩增和磁性微球，我们从没有MCS的PCR -RFLP的敏感性中浓缩了上皮细胞。 <-7>与此组合的混合在一起，我们都是门户的样本，当患者在原发性肿瘤中患有K-RAS突变时，在25％的Eventope ras the Proliphation the Oriphation Bloody中检测到该突变术后3-6个月死亡。结论和结论目前的研究表明，在患者的周围血液中存在肿瘤，每105至107个正常的核细胞可能是一个肿瘤细胞。每千兆，如果患者在外科手术前有数十种循环的癌细胞，我们认为即使是PCR-RFLP和MCS的组合也不适当地检测到胰腺癌。该系统似乎可将贡献的手术供款供应。