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Molecular detection of kidney cancers with VHL gene mutation in the blood, urine or lymph node

血液、尿液或淋巴结中 VHL 基因突变肾癌的分子检测

基本信息

批准号：
11557118
负责人：
SHUIN Taro
金额：
$ 7.17万
依托单位：
Kochi Medical School
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2001
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11557118/
关键词：
human kidney cancer VHL gene detection of cancers in the blood detection of cancers in the urine detection of cancer in the lymph node 腎細胞癌血中幡種 VHL遺伝子異常 nested RT-PCR 腫瘍マーカーリンパ節転移血中播種 nested PCR VHL癌抑制遺伝子 somatic mutation

项目摘要

According to the development of CT, MRI or ultrasonography. Human kidney cancer is detected as smaller size tumor in these days than it used to be. However, tumor metastasis to the lung, bone, brain or liver often develops more than 10 years after surgery for primary tumors. Tumor metastasis has been a major cause of death at late postoperative time. It is often stated that human kidney cancer metastatize mainly hematogeously. It is estimated that tumor cells that are living in the blood vessles is a major risk factor for tumor metastasis. We utilized tumor specific mutation in the von Hippel-Lindau disease (VHL) gene and devfeloped tumor specific PCR primer to detect cancer cells in the blood vessels just after surgery. In order to detect floating cancer cells in the blood vessels lymph node or urine in patients with kidney cancer, nested PCR were performed using specific PCR primer for specific VHL mutation. We examiined 20 patients with kidney cancer that have specific VHL mutation. We extracted RNA from the patients before during and just after surgery, at 7 days and 1 month after surgery. According to our experiment tumors specific mutation in the RNA from the blood that means tumor cells is detected more than 40 %. Tumor specific mutations are detected at a low frequency in the urine (20 %) or lymph node (5 %), respectively. It is interesting to note that tumor specific mutation is detected in the urine with single strand conformational polymorphism methods. It maskes possible to detect cancer cells before surgery without examining the site of mutation in the. We attempted to detect any tumor specific proteins that are good tumor marker. However, we could not find any promising one in the present study.Our results suggest that this molecular method for detecting cancer cell in the blood, urine or lymph node is a promising for estimating tumor invasion, or metastasis before clinical detection of kidney cancers.

根据CT、MRI或超声检查的发展。如今，人类肾癌被检测为比以前更小的肿瘤。然而，肿瘤转移至肺、骨、脑或肝的情况通常在原发肿瘤手术后 10 多年才发生。肿瘤转移已成为术后晚期死亡的主要原因。人们常说，人类肾癌主要是血行转移。据估计，生活在血管中的肿瘤细胞是肿瘤转移的主要危险因素。我们利用 von Hippel-Lindau 病 (VHL) 基因中的肿瘤特异性突变，并开发了肿瘤特异性 PCR 引物来检测手术后血管中的癌细胞。为了检测肾癌患者血管淋巴结或尿液中漂浮的癌细胞，使用针对特定VHL突变的特异性PCR引物进行巢式PCR。我们检查了 20 名具有特定 VHL 突变的肾癌患者。我们在手术前、手术中、手术后、手术后7天和1个月时从患者身上提取了RNA。根据我们的实验，肿瘤血液中 RNA 的特异性突变意味着超过 40% 的肿瘤细胞被检测到。尿液（20%）或淋巴结（5%）中分别以较低频率检测到肿瘤特异性突变。有趣的是，用单链构象多态性方法在尿液中检测到肿瘤特异性突变。它掩盖了在手术前检测癌细胞而不检查突变位点的可能性。我们试图检测任何作为良好肿瘤标志物的肿瘤特异性蛋白质。然而，我们在本研究中找不到任何有希望的方法。我们的结果表明，这种检测血液、尿液或淋巴结中癌细胞的分子方法有望在临床检测肾癌之前估计肿瘤侵袭或转移。