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Development of the new cancer cell detection method using a modified

使用改良的方法开发新的癌细胞检测方法

基本信息

批准号：
21590999
负责人：
TAKAYAMA Koichi
金额：
$ 2.83万
依托单位：
Kyushu University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2009
资助国家：
日本
起止时间：
2009 至 2011
项目状态：
已结题

项目摘要

In this study, the adenoviral vector that was modified in fiber region was used as a cancer cell detection tool. The modified adenovirus has a serotype 3 adenoviral fiber protein on the serotype 5 adenovirus, and is reported to show the high infectivity to various cancer cells in the previous paper. The purpose of this study is to improve the diagnosis accuracy for malignant pleuritis or ascites in cytological examination in combination with the modified adenoviral vector. A fire fly luciferase cDNA or GFP cDNA driven by CMV promoter are recombinated into the modified adenoviral genome using the homologous recombination method. Cancer cells infected with this recombinant modified adenoviral vectors showed a high luciferase activity or green lumination under the fluorescent microscopy. Using a diluted serum including fixed number of cancer cells as an artificial malignant effusion, the recombinant modified adenoviral vector was confirmed to detect only 10 cancer cells after RBC cells we … More re lysed out. For the clinical samples including various benign or malignant pleural effusion and ascites, the sensitivity and specificity in the diagnosis were xx% and xx%, respectively. The diagnosis accuracy with this method is a little better than that in usual cytological examination. However, in the malignant effusion with hematological malignancies like a lymphoma, the sensitivity is relatively low probably due to the lack of receptor protein for adenovirus infection on the surface of these malignant cells. On the other hand, some benign effusion also showed the higher luciferase activity resulting a pseudo-positive specimen. With a GFP expressiong modified adenoviral vector, some contaminated mesothelial cells were also infected with viral vector, and might deteriorate the diagnosis specificity. In conclusion, this method is suggested to be useful for cancer cell detection in the effusion samples in combination with cytological examination. Using a tumor specific promoter instead with CMV promoter, the diagnosis accuracy is suggested to be improved. Less

在本研究中，腺病毒载体，在纤维区被改造，被用作癌细胞检测工具。修饰后的腺病毒在血清5型腺病毒上具有血清3型腺病毒纤维蛋白，并且在之前的论文中报道对各种癌细胞表现出高感染性。本研究的目的是探讨改良腺病毒载体联合细胞学检查对恶性胸腹水的诊断准确性。使用同源重组方法将由CMV启动子驱动的萤火虫荧光素酶cDNA或GFP cDNA重组到修饰的腺病毒基因组中。用这种重组修饰的腺病毒载体感染的癌细胞在荧光显微镜下显示高荧光素酶活性或绿色发光。使用含有固定数量癌细胞的稀释血清作为人工恶性渗出液，重组修饰的腺病毒载体被证实在RBC细胞检测后仅检测到10个癌细胞。 ...更多信息再溶出。对各种良恶性胸腹水的诊断敏感性为xx%，特异性为xx%。其诊断准确率略高于常规细胞学检查。然而，在恶性血液病如淋巴瘤的恶性积液中，敏感性相对较低，可能是由于这些恶性细胞表面缺乏腺病毒感染的受体蛋白。另一方面，一些良性积液也显示较高的荧光素酶活性，导致假阳性标本。用表达GFP的腺病毒载体修饰后，一些污染的间皮细胞也被病毒载体感染，可能降低诊断的特异性。总之，这种方法是有用的渗出液样本中的癌细胞检测结合细胞学检查。用肿瘤特异性启动子代替CMV启动子，可提高诊断准确率。少