Demonstration of Biological Aggressiveness of Bone Giant Cell Tumor by the Comparative Study of Immunohistochemical Detection of DNA-instability and Cortical Bone Destruction by CT.

Abstract

The degree of DNA-instability as revealed by the immunohistochemical staining with anti-single-stranded DNA antibody after acid hydrolysis (DNA-instability test) was used as a marker of malignancy. This was applied to benign (4 osteochondroma and 4 enchondroma cases), border-line (23 bone giant cell tumor, BGCT cases), and malignant (6 chondrosarcoma and 6 osteosarcoma cases) neoplastic lesions. The expression of tumor oncogene, c-myc was detected immunohistochemically.Proliferative activity was evaluated by PCNA-immunohistochemistry, and the quantitative analysis of the number, mean area, mean total area, the largest area, and maximum shape-irregularity of AgNORs in a nucleus were performed for all these cases. The results for 19 BGCT (82.6%) cases, 6 chondrosarcoma cases (100%), and 6 osteosarcoma cases (100%) were positive with the DNA-instability test, indicating their malignancy. All benign tumor cases were negative with the DNA-instability test.Reflecting the malignant character, all chondrosarcoma cases, all osteosarcoma cases and the BGCT cases positive with the DNA-instability test showed statistically higher values of PCNA-index; and all AgNORsparameters in comparison to those for benign tumor cases, and c-myc was positive for 66.7%, and 26.3%, of them, respectively. But these values for BGCT with positive and negative DNA-instability test results showed no statistically meaningful differences. All the BGCT cases negative with DNA-instability test were negative for cmyc expression.Among the BGCT cases positive with DNA-instability test, 18 cases (94.7%) showed cortical bone destruction by computed tomography (CT), and 5 cases (26.3%) showed extra-osseous expansion. No such radiographic changes were detected among the BGCT cases negative with DNAinstability test. Among 18 BGCT cases with cortical bone destruction, 5 cases (27.8%) showed tumor recurrences, and 2 cases (11.1 %) showed lung metastases.These results indicate that the majority of BGCT cases are malignant and the detection of cortical bone destruction by CT is a sensitive clinical marker to detect them.

Detection of cancer clones in human gastric adenoma by increased DNA-instability and other biomarkers

“…In these studies, and also in the present study, the DNA-instability-positive lesions showed significantly higher values of positivity of other biomarkers. Furthermore, many abnormal mitoses were found predominantly in the DNA-instability test-positive lesions as shown in the present study (Figure 2) and other reports (Tsuzuki et al, 1994;Azuchi et al, 1998); and the frequency of chromosome 17 polysomies and the percentage of hyperdiploid cells measured by means of the chromosome index using interphase cytogenetics increased significantly and exclusively in the DNA-instability test-positive lesions (Khaled et al, 2000;Ishida et al, 2001). This is also supported by the report that numerical chromosomal aberration is associated with cancer progression (Bulten et al, 1998).…”

“…Based on the finding that all cancerous cells produce much more single-stranded DNA than normal cells after acid hydrolysis, Fukuda et al (1986Fukuda et al ( , 1993 developed the method of staining cancerous cells specifically with acridine orange or anti-cytidine antibody after acid hydrolysis and called it as the DNA-instability test as a specific marker for malignancy irrespective of epithelial or mesenchymal origin. Since the first report (Fukuda et al, 1986), more than 20,000 malignancy cases and socalled borderline malignancy cases were tested by this method (Nitta et al, 1993;Otaki et al, 1994;Tsuzuki et al, 1994;Azuchi et al, 1998;Khaled et al, 2000;Hirai et al, 2001;Ishida et al, 2001;Iwasa et al, 2001), and all cancerous cells were found to be positively stained in the DNA-instability test, while comparable normal cells, vascular endothelial cells, fibroblasts and inflammatory cells present in the same slides were completely negative.…”

“…They also developed the method of differential fluorescent or immunohistochemical staining of cancerous cells after denaturation of DNA by acid hydrolysis. Using this method (the DNA-instability test), cancerous clones could be identified during early stages of carcinogenesis in so-called borderline or precancerous lesions such as colorectal ade-noma (Nitta et al, 1993), otorhinolaryngeal borderline lesions (Tsuzuki et al, 1994), bone giant cell tumors (Azuchi et al, 1998), uterine cervical dysplasia (Khaled et al, 2000), oral leukoplakia (Iwasa et al, 2001), 20-methylcholanthreneinduced squamous cell carcinoma of mouse epidermis (Hirai et al,2001). Otaki et al (1994) also demonstrated that all adenoma lesions with severe dysplasia can be regarded as malignant in nature existing at an early stage of cancer progression, using the DNA-instability test compared with PCNA immunohistochemistry and AgNORs parameter analysis.…”

Immunohistochemical detection of early-stage carcinogenesis of oral leukoplakia by increased DNA-instability and various malignancy markers

“…The DNA-instability-test has been proposed by us as a specific marker of malignancy, irrespective of the epithelial or mesenchymal origin of tumors (Fukuda et al, 1986(Fukuda et al, , 1993Nitta et al, 1993;Otaki et al, 1994;Tsuzuki et al, 1994;Azuchi et al, 1998;Khaled et al, 2000). The method of differentially staining malignant cells either with acridine orange or anti-single-stranded DNA is based on the abundant production of denatured singlestanded DNA in cancerous cells after acid hydrolysis.…”

“…They also developed the method of differential fluorescent or immunohistochemical staining of cancerous cells after denaturation of nuclear DNA by acid hydrolysis. By using this method (the DNA instability test), cancerous clones could be identified during early stages of carcinogenesis in so-called borderline or precancerous lesions such as colorectal adenoma (Nitta et al, 1993), gastric adenoma (Otaki et al, 1994), otorhinolaryngeal borderline lesions (Tsuzuki et al, 1994), bone giant cell tumours (Azuchi et al, 1998), and uterine cervical dysplasia (Khaled et al, 2000).…”