ABSTRACT
The specimens of ductal carcinoma in situ (DCIS) with early invasion, and specimens collected by core needle biopsy (CNB) tend to contain limited amount of invasive component, so it is imperative to explore a new technique which can assess HER2 gene status accurately for the limited invasive cancer component in these specimens. Dual staining technique of combining immunohistochemistry (IHC) for myoepithelial cells and single or dual probe chromogenic in situ hybridization (CISH) for HER2 gene was performed on routinely processed paraffin sections from 20 cases diagnosed as having DCIS with invasive cancer. Among them, 10 had fluorescence in situ hybridization (FISH)-confirmed amplification of HER2 and 10 had FISH-confirmed non-amplification of HER2. We successfully detected HER2 genetic signals and myoepithelial IHC markers (SMM-HC or CK5/6) simultaneously on a single section in all 20 specimens. Myoepithelial markers and HER2 signals detected by dual staining assay were consistent with those by individual technique performed alone. HER2 gene amplification results determined by dual staining assay were 100% consistent with those of FISH. Dual staining technique which allows simultaneous detection of myoepithelial marker protein and cancerous HER2 gene is feasible, and it has potential to be used in clinical practice for effective determination of HER2 amplification in limited invasive component.
ABSTRACT
The specimens of ductal carcinoma in situ (DCIS) with early invasion, and specimens collected by core needle biopsy (CNB) tend to contain limited amount of invasive component, so it is imperative to explore a new technique which can assess HER2 gene status accurately for the limited invasive cancer component in these specimens. Dual staining technique of combining immunohistochemistry (IHC) for myoepithelial cells and single or dual probe chromogenic in situ hybridization (CISH) for HER2 gene was performed on routinely processed paraffin sections from 20 cases diagnosed as having DCIS with invasive cancer. Among them, 10 had fluorescence in situ hybridization (FISH)-confirmed amplification of HER2 and 10 had FISH-confirmed non-amplification of HER2. We successfully detected HER2 genetic signals and myoepithelial IHC markers (SMM-HC or CK5/6) simultaneously on a single section in all 20 specimens. Myoepithelial markers and HER2 signals detected by dual staining assay were consistent with those by individual technique performed alone. HER2 gene amplification results determined by dual staining assay were 100% consistent with those of FISH. Dual staining technique which allows simultaneous detection of myoepithelial marker protein and cancerous HER2 gene is feasible, and it has potential to be used in clinical practice for effective determination of HER2 amplification in limited invasive component.
Subject(s)
Female , Humans , Biomarkers, Tumor , Metabolism , Breast Neoplasms , Genetics , Metabolism , Pathology , Chromogenic Compounds , Gene Expression Profiling , Methods , Immunohistochemistry , Methods , In Situ Hybridization, Fluorescence , Methods , Neoplasm Invasiveness , Pathology , Receptor, ErbB-2 , Genetics , Metabolism , Reproducibility of Results , Sensitivity and SpecificityABSTRACT
Gastric carcinoma with osteoclast-like giant cells (OGCs) is an extremely rare tumor. So far, only six cases have been reported in the literature. Here we report an additional case of this tumor in a Chinese 78-year-old man presented with abdominal pain, vomiting, and hematemesis. Physical examination and gastroscopy revealed a tumor in the gastric antrum. The biopsy and pathological findings indicated a gastric adenocarcinoma with OGCs, which were present in both the tumor and the metastatic lymph nodes. Further immunohistochemical staining indicated that OGCs were reactive with CD68, CD45, and vimentin protein, but not with pancytokeratin, carcinoembryonic antigen, or epithelial membrane antigen, suggesting the monocytic/histiocytic derivation of these OGCs. In situ hybridization for Epstein-Barr virus showed no nuclear positivity in either adenocarcinoma or OGCs. Postoperative follow-up showed that the patient had survived for at least 6 months without recurrence. Further investigation is warranted to clearly define the prognostic significance of OGCs in gastric carcinoma.
Subject(s)
Aged , Humans , Male , Giant Cells , Metabolism , Pathology , Immunohistochemistry , In Situ Hybridization , Osteoclasts , Metabolism , Pathology , Stomach Neoplasms , Genetics , Metabolism , PathologyABSTRACT
<p><b>OBJECTIVE</b>To detect tumor specific chromosome translocations and associated fusion transcripts in paraffin-embedded tissue by interphase fluorescence in-situ hybridization (FISH) and reverse transcription polymerase chain reaction (RT-PCR), respectively, and to evaluate their diagnostic values for Ewing's sarcoma/primitive neuroectodermal tumor (ES/PNET).</p><p><b>METHODS</b>Nuclei of the tumor cells and total RNA were extracted from 10 cases of ES/PNET. Interphase FISH was utilized to analyze the EWS gene translocation with a dual color, break apart probe (Vysis company). RT-PCR was used to detect t (11; 22) (q24; q12) and t (21; 22) (q22; q12) fusion transcripts.</p><p><b>RESULTS</b>Among 10 cases of ES/PNET, the EWS-FLI1 fusion transcript was detected in 8 by RT-PCR. EWS-ERG fusion transcript was not detected in any of the cases. EWS gene translocation was found in 9 of 10 cases by FISH.</p><p><b>CONCLUSIONS</b>Interphase FISH and RT-PCR can be reliably applied to paraffin-embedded tissues for molecular diagnosis of ES/PNET. Between the two approaches, interphase FISH provides a more sensitive and stable result.</p>
Subject(s)
Adolescent , Adult , Child , Humans , In Situ Hybridization, Fluorescence , Methods , Neuroectodermal Tumors, Primitive, Peripheral , Diagnosis , Genetics , Polymerase Chain Reaction , Reverse Transcriptase Polymerase Chain Reaction , Methods , Sarcoma, Ewing , Diagnosis , GeneticsABSTRACT
<p><b>OBJECTIVES</b>To study the expression and the significance of telomerase gene hTERT in testes of infertile male.</p><p><b>METHODS</b>By using in situ hybridization(ISH) techniques, the expression of telomerase gene hTERT mRNA in testes of 47 infertile male and 10 normal testicular tissues were observed.</p><p><b>RESULTS</b>In male testes, there was a positive correlation between the expression of hTERT and the quantity and density of germ cells(spermatogonia, spermatocyte, spermatid). The expression of hTERT in some germinal cell of maturation arrest patients were not significantly different with those of normal.</p><p><b>CONCLUSIONS</b>Our results suggest that the deficiency of telomerase might be a factor for germinal cell maturation arrest and there might be some other etiological factors in these patients. Our study provides experimental groundwork for the gene therapy of male infertility.</p>