Showing papers by "Nam Jin Yoo published in 1998"

A simple, precise and economical microdissection technique for analysis of genomic DNA from archival tissue sections

Abstract: Formalin-fixed and paraffin-embedded tissues are valuable resources for retrospective analysis of the molecular changes in DNA present in tumour tissues. One common problem that precludes an accurate DNA analysis in a human tissue sample is cellular heterogeneity. We have developed a simple and inexpensive, but micrometrically precise, microdissection technique that allows for selective isolation of minute cell clusters and even single cells from archival tissue sections. The features of our technique include use of a 30G1/2 needle affixed to a mechanical micromanipulator as a dissector sharp enough to be used for dissection of even single cells and use of the stage and focus control knobs of the microscope to scrape the target cells instead of moving the needle during microdissection. The main advantages of this technique over the current methods lie in its simplicity, low cost, easy handling and precision.

Loss of Heterozygosity and Somatic Mutations of the VHL Tumor Suppressor Gene in Sporadic Cerebellar Hemangioblastomas

Abstract: Cerebellar hemangioblastoma is a benign central nervous system neoplasm with characteristic proliferation of vascular and stromal cells. There is increasing evidence that the stromal cell population may represent the neoplastic component of hemangioblastoma, whereas the vascular component may be composed of reactive, nonneoplastic cells. Therefore, successful genetic testing for loss of heterozygosity requires selective analysis of target cell populations. Here, tissue microdissection was used to selectively analyze the stromal cell component of 20 archival sporadic cerebellar hemangioblastomas for loss of heterozygosity at the Von-Hippel Lindau (VHL) gene and somatic VHL gene mutations. Allelic deletions at the VHL gene locus were detected in the stromal cell component with one or more markers (D3S1038, D3S1110, and/or 104/105) in 10 of 19 (52.6%) informative cases. In all cases, heterozygosity at the VHL gene locus was retained in the vascular component. In two cases, aberrant bands in exon 2 of the VHL gene were demonstrated in the stromal cells by PCR-based single-strand conformation polymorphism analysis, and somatic missense mutations were successfully characterized in two of the sporadic hemangioblastomas by direct sequencing. The results suggest that allelic losses and mutations of the VHL tumor suppressor gene play a role in sporadic cerebellar hemangioblastoma tumorigenesis. Furthermore, because the genetic changes were detected in selectively procured stromal cell areas, the data provide strong evidence that the stromal cell represents a neoplastic component of hemangioblastoma.

Frequent Somatic Mutations in Serine/Threonine Kinase 11/Peutz-Jeghers Syndrome Gene in Left-sided Colon Cancer

Abstract: We analyzed somatic mutation and loss of heterozygosity (LOH) in the serine/threonine kinase 11 (STK11)/Peutz-Jeghers syndrome gene in 49 colorectal tumors in three different stages of a dysplasia-carcinoma sequence. We detected LOH in 10 of 19 (52.6%) informative colorectal cancers at loci D19S886 and/or D19S883, but no LOH was observed in 25 informative adenomas. We detected a total of 9 somatic mutations [7 of 13 (53.8%) left-sided colon cancers and 2 of 7 (28.6%) left-sided adenomas with high-grade dysplasia], but no mutations were detected in right-sided colon tumors. Of the nine mutations, one was a frameshift mutation (the same mutation detected in Peutz-Jeghers syndrome family previously), and the other eight were missense mutations. This results indicate that STK11 is a tumor suppressor gene and that genetic changes of STK11 play an important role in left-sided colon cancer carcinogenesis.

A distinct region of chromosome 19p13.3 associated with the sporadic form of adenoma malignum of the uterine cervix

Abstract: Adenoma malignum (AM) is known to be one of the malignant tumors that is commonly associated with Peutz-Jeghers syndrome. Recently, the genetic locus of Peutz-Jeghers syndrome was mapped to the telomeric region of chromosome 19p. We analyzed nine sporadic cases of AM with high-density loss of heterozygosity to study the region of chromosome 19p13.2-13.3 using eight microsatellite markers. Our deletion mapping data revealed a distinct region with 100% loss of heterozygosity frequency at marker D19S216. This result indicates that a putative tumor suppressor gene for AM is located at D19S216 on chromosomal band 19p13.3 and plays an important role in AM tumorigenesis.

Immunohistochemical analysis of Fas ligand expression in sarcomas. Sarcomas express high level of FasL in vivo

Abstract: Fas ligand (FasL) and its receptor, Fas, play a key role in the regulation of apoptosis within the immune system. Several prior experimental studies of Fas ligand expression in tumors have suggested a mechanism that enables tumors to evade immune destruction by inducing apoptosis in activated lymphocytes near the tumor cells. Many types of carcinomas have been shown to express FasL, but at present nothing is known about the range of sarcomas capable of expressing FasL in vivo. The aim of this study was to determine the in vivo patterns of FasL expression in human sarcomas. Archival paraffin-embedded tissues of 57 sarcomas and 30 carcinomas were analyzed by immunohistochemistry for the expression of FasL. FasL immunoreactivity was seen in 39 of 57 (68%) sarcomas, including 10 of 10 rhabdomyosarcomas, 5 of 5 malignant schwannomas, 2 of 2 Ewing's sarcomas, 8 of 11 malignant fibrous histiocytomas, 4 of 5 angiosarcomas, 2 of 5 synovial sarcomas, 2 of 5 liposarcomas, 3 of 5 leiomyosarcomas, 2 of 6 osteosarcomas, and 1 of 3 chondrosarcomas. All carcinomas tested (10 gastric adenocarcinomas, 10 hepatocellular carcinomas, and 10 renal cell carcinomas) were positive for FasL. These results demonstrate that FasL expression in sarcomas, although less frequent than in carcinomas, is widespread among the sarcoma types, and suggest that FasL might contribute to the immune escape of sarcomas through killing Fas-bearing lymphocytes.