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

根据CT、MRI或超声检查的发展。人类肾癌在这些天被检测为比过去更小的肿瘤。然而，肿瘤转移到肺、骨、脑或肝通常在原发性肿瘤手术后超过10年发展。肿瘤转移是术后晚期死亡的主要原因。通常认为人肾癌主要通过血液转移。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VHL基因的肿瘤特异性突变，研制了肿瘤特异性PCR引物，用于检测手术后血管中的癌细胞。为检测肾癌患者血管、淋巴结或尿液中漂浮的癌细胞，采用特异性VHL突变的PCR引物进行巢式PCR。我们检查了20例具有特异性VHL突变的肾癌患者。我们在手术前、手术中、手术后第7天和手术后1个月从患者中提取RNA。根据我们的实验，肿瘤特异性突变的RNA来自血液，这意味着肿瘤细胞被检测到超过40%。在尿液（20%）或淋巴结（5%）中分别检测到低频率的肿瘤特异性突变。值得注意的是，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。我们试图检测任何肿瘤特异性蛋白，这是良好的肿瘤标志物。本研究的结果提示，这种从血液、尿液或淋巴结中检测癌细胞的分子生物学方法有希望在临床检测肾癌前评估肿瘤的侵袭或转移。

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.