No abstract available.
Fluorescence* ; In Situ Hybridization*

A 35-year-old man with infertility was referred for chromosomal analysis. In routine cytogenetic analysis, the patient was seen to have additional material of unknown origin on the terminal region of the short arm of chromosome 4. To determine the origin of the unknown material, we carried out high-resolution banding, comparative genomic hybridization (CGH), and FISH. CGH showed a gain of signal on the region of 4q32-->q35. FISH using whole chromosome painting and subtelomeric region probes for chromosome 4 confirmed the aberrant chromosome as an intrachromosomal insertion duplication of 4q32-->q35. Duplication often leads to some phenotypic abnormalities; however, our patient showed an almost normal phenotype except for congenital dysfunction in spermatogenesis.
Adult ; Arm ; Chromosome Painting ; Chromosomes, Human, Pair 4 ; Comparative Genomic Hybridization ; Cytogenetic Analysis ; Humans ; In Situ Hybridization, Fluorescence ; Infertility ; Phenotype ; Spermatogenesis ; Trisomy

No abstract available.
Humans ; In Situ Hybridization, Fluorescence ; Infant, Newborn

No abstract available.
Humans ; In Situ Hybridization, Fluorescence ; Infant, Newborn

PURPOSE: The recent discovery and characterization of an oncogenic ROS1 gene rearrangement has raised significant interest because small molecule inhibitors are effective in these tumors. The aim of this study was to determine frequency and clinicopathological features associated with ROS1 rearrangement in patients with cholangiocarcinoma (CCA). MATERIALS AND METHODS: A total of 261 patients who underwent surgery for CCA between October 1997 and August 2013 were identified from an international, multi-institutional database. ROS1 rearrangement was evaluated by break-apart fluorescence in situ hybridization using tissue microarrays of these patients. RESULTS: Of 261 CCA evaluated, three cases (1.1%) showed ROS1 rearrangement by fluorescence in situ hybridization (FISH), all of which were derived from intrahepatic origin. ROS1 protein expression was observed in 38 samples (19.1%). Significantly larger tumor size was observed in ROS1 immunohistochemistry (IHC)–negative patients compared with ROS1 IHC–positive patients. ROS1 FISH–positive patients had a single tumor with a median size of 4 cm and well-to-moderate differentiation. Overall, there was no difference in terms of baseline characteristics, overall survival, and recurrence-free survival between ROS1-positive and -negative patients. CONCLUSION: ROS1 rearrangement was detected in 1.1% of CCA patients. Although rare, conduct of clinical trials using ROS1 inhibitors in these genetically unique patients is warranted.
Cholangiocarcinoma* ; Fluorescence ; Gene Rearrangement ; Humans ; Immunohistochemistry ; In Situ Hybridization ; In Situ Hybridization, Fluorescence ; Incidence*

BACKGROUND: The amplification of murine double minutes (MDM2) is the primary feature of well-differentiated liposarcomas (WDLPS) and dedifferentiated liposarcomas (DDLPS), while DDIT3 rearrangement is the main one of myxoid liposarcomas (MLPS). Our aim was to evaluate the added value of MDM2 amplification and DDIT3 rearrangement in making a diagnosis and classifying lipogenic tumors. METHODS: Eighty-two cases of liposarcoma and 60 lipomas diagnosed between 1995 and 2010 were analysed for MDM2 amplification and DDIT3 rearrangement using a fluorescence in situ hybridization (FISH). The subtypes of liposarcoma were reclassified according to the molecular results, whose results were reviewed with an analysis of the relevant histologic and immunohistochemical findings. RESULTS: One case of lipoma (1.67%) was reclassified as a WDLPS. Of the liposarcomas, 13.4% (16/82) were reclassified after the molecular testing. Five cases of MLPS were reclassified as four cases of DDLPS and one case of myxoid lipoma. Two cases of WDLPS were reclassified as one case of spindle cell lipoma and another case of myxofibrosarcoma. Four cases of DDLPS were reclassified as two cases of leiomyosarcoma, one case of angiomyolipoma and another case of fibroinflammatory lesion. Of the six cases of pleomorphic liposarcoma, five were reclassified as DDLPS. CONCLUSIONS: In our series, a critical revision of diagnosis was found at a rate of 3.5% (5/142) after a review of the lipomatous lesions. The uses of molecular testing by MDM2 and DDIT3 FISH were valuable to make an accurate subtyping of liposarcomas as well as to differentiate WDLPS from benign lipomatous tumor.
Angiomyolipoma ; Fluorescence ; In Situ Hybridization ; In Situ Hybridization, Fluorescence ; Leiomyosarcoma ; Lipoma ; Liposarcoma ; Liposarcoma, Myxoid

PURPOSE: The characterization of all recognizable chromosomal rearrangements was dis- turbed by technical limitation of conventional cytogenetic methods. Recently, the strong usefullness of generation of chromosome specific painting probes in identification of marker chromosomes has proven. This study was intended to analyze the chromosomal aberrations in human ovarian cancer cell line, SNU-8, by G-banding and multiple paintings. MATERIALS AND METHODS: Human ovarian cancer cell line, SNU-8 was cultured and harvested for cytogenetic analysis. Routine karyotyping was performed. For complete analysis of chromosomal aberrations, human chromosome-specific painting probes were constructed from somatic hybrid cells. The origins of the unidentified marker chromosomes were analyzed by fluorescent in situ hybridization (FISH) with these painting probes. RESULTS: All chromosome alterations were confirmed by the use of multiple chromosome paintings, which also demonstrated a number of additional alterations. SNU-8 had the karyotype 62-69,XXX, + der(1;10)(q10;p10),der(3;18) (q10;p10)X2,-4,+ 5,+ 7,del(9)(q21)X2,-11,-13,-15,-16,der(17;19)(q10;q10) X2, + 20,-22[cp51]. CONCLUSION: The chromosomal aberrations of SNU-8 cell line was effectively analyzed by FISH with these painting probes, and the approach methods of this study can be applied to cytogenetic analysis of chromosomal aberrations in the other cancers.
Cell Line ; Cell Line, Tumor ; Chromosome Aberrations ; Chromosome Painting ; Cytogenetic Analysis ; Cytogenetics ; Humans ; Hybrid Cells ; In Situ Hybridization, Fluorescence ; Karyotype ; Karyotyping ; Ovarian Neoplasms ; Paint ; Paintings

PURPOSE: There are only a few cytogenetic studies in gastric cancer and so far no consistent specific chromosomal abnormalities have been described. In this study, we have used comparative genomic hybridization (CGH), a powerful molecular cytogenetic technique for detecting changes of the copy number throughout the genome, to screen for genetic alterations in gastric cancer cell lines. The CGH results were compared with those derived from G-banding and chromosome painting. MATERIALS AND METHODS: Conventional cytogenetic analysis was performed on five human gastric cancer cell lines, AGS, SNU-1, SNU-16, SNU-620, and SNU-719, by a G-banding staining technique. In CGH, equal amounts of differently labeled DNA from the cell lines and normal reference DNA were hybridized simultaneously to normal metaphase chromosomes. They were visualized by different fluorochromes, and the signal intensities were quantitated separately as gray levels along the single chromosomes. The over- and under- represented DNA segments were determined by computation of ratio images and average ratio profiles. To confirm the CGH results, fluorescence in situ hybridization (FISH) with chromosome specific painting was performed using indirectly labeled chromosome specific paints. RESULTS: Complex unbalanced chromosomal aberrations that could not be identified reliably by conventional cytogenetics in gastric cancer cell lines were successfully resolved by CGH analysis. CGH results were validated by using FISH with chromosome specific probes. In gastric cancer cell lines, gains of DNA copy number were more common than losses. Gains were detected on 1p, 1q, 2p, 3q, 6p, 7q, 10q, 11p, and 19q, and losses were observed on 4p, 4q, 5q, 12p, 12q, and 18q. Interestingly, all the five gastric cancer cell lines tested showed gain of DNA copy number on the chromosome 20, suggesting an existence of oncogene. CONCLUSION: Conventional cytogenetics, CGH, and FISH using painting probes represent complementary approaches that, when employed in combination, could greatly facilitate the comprehensive analysis of chromosomal imbalances in gastric cancer cell lines. Our results suggest the existence of an oncogene or oncogenes on chromosome 20 that play a role in the development and/or the progression of gastric carcinogenesis.
Carcinogenesis ; Cell Line ; Chromosome Aberrations ; Chromosome Painting ; Chromosomes, Human, Pair 20 ; Comparative Genomic Hybridization ; Cytogenetic Analysis ; Cytogenetics ; DNA ; Fluorescence ; Fluorescent Dyes ; Genome ; Humans ; In Situ Hybridization ; Metaphase ; Oncogenes ; Paint ; Paintings ; Stomach Neoplasms

PURPOSE: There are only a few cytogenetic studies in gastric cancer and so far no consistent specific chromosomal abnormalities have been described. In this study, we have used comparative genomic hybridization (CGH), a powerful molecular cytogenetic technique for detecting changes of the copy number throughout the genome, to screen for genetic alterations in gastric cancer cell lines. The CGH results were compared with those derived from G-banding and chromosome painting. MATERIALS AND METHODS: Conventional cytogenetic analysis was performed on five human gastric cancer cell lines, AGS, SNU-1, SNU-16, SNU-620, and SNU-719, by a G-banding staining technique. In CGH, equal amounts of differently labeled DNA from the cell lines and normal reference DNA were hybridized simultaneously to normal metaphase chromosomes. They were visualized by different fluorochromes, and the signal intensities were quantitated separately as gray levels along the single chromosomes. The over- and under- represented DNA segments were determined by computation of ratio images and average ratio profiles. To confirm the CGH results, fluorescence in situ hybridization (FISH) with chromosome specific painting was performed using indirectly labeled chromosome specific paints. RESULTS: Complex unbalanced chromosomal aberrations that could not be identified reliably by conventional cytogenetics in gastric cancer cell lines were successfully resolved by CGH analysis. CGH results were validated by using FISH with chromosome specific probes. In gastric cancer cell lines, gains of DNA copy number were more common than losses. Gains were detected on 1p, 1q, 2p, 3q, 6p, 7q, 10q, 11p, and 19q, and losses were observed on 4p, 4q, 5q, 12p, 12q, and 18q. Interestingly, all the five gastric cancer cell lines tested showed gain of DNA copy number on the chromosome 20, suggesting an existence of oncogene. CONCLUSION: Conventional cytogenetics, CGH, and FISH using painting probes represent complementary approaches that, when employed in combination, could greatly facilitate the comprehensive analysis of chromosomal imbalances in gastric cancer cell lines. Our results suggest the existence of an oncogene or oncogenes on chromosome 20 that play a role in the development and/or the progression of gastric carcinogenesis.
Carcinogenesis ; Cell Line ; Chromosome Aberrations ; Chromosome Painting ; Chromosomes, Human, Pair 20 ; Comparative Genomic Hybridization ; Cytogenetic Analysis ; Cytogenetics ; DNA ; Fluorescence ; Fluorescent Dyes ; Genome ; Humans ; In Situ Hybridization ; Metaphase ; Oncogenes ; Paint ; Paintings ; Stomach Neoplasms

PURPOSE: Most tests developed for the determination of the HER2 status still require validation, although identification of the HER2 status is important for predicting the response to specific systemic therapy in breast cancer. Fluorescence in situ hybridization (FISH) and immunohistochemistry (IHC) were performed for the HER2 expression on the tissue microarray (TMA) from primary breast cancers to validate the feasibility of IHC for HER2 assay. METHODS: A TMA was constructed from 134~231 primary breast cancers. FISH and IHC were repeated more than twice, and the results were analyzed. Three kinds of primary antibody for IHC were used and compared. RESULTS: The HER2 was amplified by FISH in 24~28% of breast cancer with a concordance between multiple assays of 92~100% (kappa=0.994-0.965), while the HER2 was overexpressed in 21~27% by IHC. The HER2 was amplified in 70~100% of the IHC 3+ cases, but was observed in only 45~78% and 5~12% of the IHC 2+ and IHC 0~1+ cases, respectively. The results from the IHC, using 3 different primary antibodies to HER2, were in good agreement each other at 88~92% (kappa=0.902-0.799). CONCLUSION: The results of the FISH appeared to be more reproducible than those of the IHC in the current study. The results of the IHC were not different from each other according to primary antibody used. However, a considerable proportions of the IHC positive cases were not confirmed by the FISH. This report indicates a need to improve the laboratory quality control measures when using the IHC for the HER2 assay, including the periodic testing for the concordance with FISH.
Antibodies ; Breast Neoplasms* ; Breast* ; Fluorescence* ; Immunohistochemistry* ; In Situ Hybridization* ; Quality Control