Development of the new cancer cell detection method using a modified

本研究利用在纤维区修饰的腺病毒载体作为癌细胞检测工具。修饰后的腺病毒在血清5型腺病毒上有一个血清3型腺病毒纤维蛋白，文献报道对多种癌细胞具有较高的感染性。本研究旨在结合改良的腺病毒载体，提高细胞学检查对恶性胸膜炎或腹水的诊断准确性。利用同源重组法将CMV启动子驱动的荧光素酶cDNA或GFP cDNA重组到修饰的腺病毒基因组中。经重组修饰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。用含有固定数量癌细胞的稀释血清作为人工恶性积液，证实重组修饰腺病毒载体在红细胞溶解后仅能检测到10个癌细胞。对于包括各种良恶性胸腔积液和腹水在内的临床标本，诊断的敏感性和特异性分别为xx%和xx%。该方法的诊断准确率略高于常规细胞学检查。然而，在血液系统恶性肿瘤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。使用表达GFP修饰的腺病毒载体，一些污染的间皮细胞也被病毒载体感染，可能会降低诊断特异性。本方法与细胞学检查相结合，可用于积液标本的癌细胞检测。用肿瘤特异性启动子代替CMV启动子可提高诊断准确性。少

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