• Title, Summary, Keyword: silver in situ hybridization

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Determination of HER2 Gene Amplification in Breast Cancer using Dual-color Silver Enhanced in situ Hybridization (dc-SISH) and Comparison with Fluorescence ISH (FISH)

  • Unal, Betul;Karaveli, Fatma Seyda;Pestereli, Hadice Elif;Erdogan, Gulgun
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.10
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    • pp.6131-6134
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    • 2013
  • Background: The two basic methods that are currently accepted to identify the HER2 status are immunohistochemistry and flyorescence in situ hybridization (FISH). The aim of this study was to perform the dual-color silver in situ hybridization (dc-SISH) technique as an alternative to FISH. Materials and Methods: A total of 40 invasive breast carcinoma cases were assessed for HER2 gene amplification by FISH and dual-color SISH. Results: Significant correlation was found in the HER2 expression results obtained with the two approaches (p=0.001, p<0.05). The concordance rate was 92.3%. Conclusions: Foutine practical use of the dc-SISH method, which is much easier to apply, score, and evaluate, has many advantages. HER2 and CEN17 status can be evaluated simultaneously with the newly developed "Dual-Color Probe". All these specifications and the reliable results obtained support the widespread use of SISH technique in clinical practice.

Detection of Pneumocvstis carinii by in situ hybridization in the lungs of immunosuppressed rats (면역억제 흰쥐에서 조직내교잡법을 이용한 페포자충의 검출)

  • Jin KIM;Jae-Ran YU;Sung-Tae HONG;Chang-Soo PARK
    • The Korean Journal of Parasitology
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    • v.34 no.3
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    • pp.177-184
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    • 1996
  • In situ hybridization was performed to detect rat heumocwstis ca4nii in the lung sections. Rats were immunosuppressed by weekly subcutaneous injection of 10 mg/kg methylprednisolone. On the 6th, 8th and 9th week of immunosuppression, the lungs were removed and fled in 10% neutral formalin. A 22 base oligonucleotide probe complementary to p. carinii 5S ribosomal RMh was commercially synhesized and its 3' terminal was labeled wiH biotin. In situ hybridization was performed utilizing manual capillary action technolog)r on the Microprobe system. p. cnrinii were detected along the luminal surface of alveolar pneumocytes, in exudate of alveolar cavities, and also in secretory material of bronchioles. In the 6th week group, positive reaction was observed focally in the peripheral region of the lung sections, but the reaction was observed diffusely in the 8th or 9th week groups. In comparison with Grocott's methenamine silver stain, in situ hybridization technique can detect the organism rapidly, and can detect trophic forms very well. Furthermore, no nonspecific reaction with other pathogenic fungi and protozoa was recognized. Therefore, in situ hybridization can be a good technique to detect p. carinii in the lungs of infected rats.

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Identification of Epstein-Barr Virus in the Human Placenta and Its Pathologic Characteristics

  • Kim, Younghoon;Kim, Hye Sung;Park, Joong Shin;Kim, Chong Jai;Kim, Woo Ho
    • Journal of Korean Medical Science
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    • v.32 no.12
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    • pp.1959-1966
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    • 2017
  • Epstein-Barr virus (EBV), a common pathogen in humans, is suspected as the cause of multiple pregnancy-related pathologies including depression, preeclampsia, and stillbirth. Moreover, transmission of EBV through the placenta has been reported. However, the focus of EBV infection within the placenta has remained unknown to date. In this study, we proved the expression of latent EBV genes in the endometrial glandular epithelial cells of the placenta and investigated the cytological characteristics of these cells. Sixty-eight placentas were obtained from pregnant women. Tissue microarray was constructed. EBV latent genes including EBV-encoding RNA-1 (EBER1), Epstein-Barr virus nuclear antigen 1 (EBNA1), late membrane antigen (LMP1), and RPMS1 were detected with silver in situ hybridization and/or mRNA in situ hybridization. Nuclear features of EBV-positive cells in EBV-infected placenta were compared with those of EBV-negative cells via image analysis. Sixteen placentas (23.5%) showed positive expression of all 4 EBV latent genes; only the glandular epithelial cells of the decidua showed EBV gene expression. EBV infection status was not significantly correlated with maternal, fetal, or placental factors. The nuclei of EBV-positive cells were significantly larger, longer, and round-shaped than those of EBV-negative cells regardless of EBV-infection status of the placenta. For the first time, evidence of EBV gene expression has been shown in placental tissues. Furthermore, we have characterized its cytological features, allowing screening of EBV infection through microscopic examination.

Is Her-2 Status in the Primary Tumor Correlated with Matched Lymph Node Metastases in Patients with Gastric Cancer Undergoing Curative Gastrectomy?

  • Selcukbiricik, Fatih;Erdamar, Sibel;Buyukunal, Evin;Serrdengecti, Suheyla;Demirelli, Fuat
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.24
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    • pp.10607-10611
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    • 2015
  • Background: HER2 expression in the primary tumor and its lymph node metastases vary in gastric cancer, reflecting intratumoral heterogeneity. This finding also suggests that proliferation of a different clone in metastatic nodes is possible. In the current study, we aimed to determine the cause of discordance in HER-2 expression in the primary tumor and lymph node metastases for patients with gastric cancer. Materials and Methods: Eighty-one patients with gastric cancer who had undergone radical gastrectomy and were found to have lymph node metastasis upon pathological examination were included. Histopathological samples were obtained from biopsies obtained during patient gastrectomies and lymph node dissection. HER2 status was evaluated by both immunohistochemistry (IHC) and silver in situ hybridization (SISH). Results: Sixty-four (79%) patients were SISH (-), while 17 (21%) were SISH (+) in the primary tumor. However, in metastatic lymph nodes, HER2 status was SISH positive in 5 (28.3%) of the 64 SISH (-) primary tumor specimens. One of the 17 SISH (+) primary tumors was SISH (-) in the metastatic lymph nodes. Thus, SISH results for HER2 in both primary tumors and lymph node metastases were comparable, showing a concordance of 92.5%. In total, six patients demonstrated discordance between the primary tumor and lymph node metastases. The prevalence of HER2 discordance was significantly higher for patients in the pN2 and N3 stages (p=0.007). Although discordant patients had worse survival rates than concordant patients, the differences were not significant (p>0.05).Conclusions: Our study indicates that the frequency of concordance in HER2 status, as determined by IHC or SISH, is high in primary tumors and their corresponding lymph node metastases for patients with gastric cancer. If there is a discrepancy in HER2 status, its evaluation by both IHC and SISH may be useful for detecting patients who would benefit from trastuzumab, and it would therefore help guide decision-making processes in administering treatment.

Highly Sensitive Biological Analysis Using Optical Microfluidic Sensor

  • Lee, Sang-Yeop;Chen, Ling-Xin;Choo, Jae-Bum;Lee, Eun-Kyu;Lee, Sang-Hoon
    • Journal of the Optical Society of Korea
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    • v.10 no.3
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    • pp.130-142
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    • 2006
  • Lab-on-a-chip technology is attracting great interest because the miniaturization of reaction systems offers practical advantages over classical bench-top chemical systems. Rapid mixing of the fluids flowing through a microchannel is very important for various applications of microfluidic systems. In addition, highly sensitive on-chip detection techniques are essential for the in situ monitoring of chemical reactions because the detection volume in a channel is extremely small. Recently, a confocal surface enhanced Raman spectroscopic (SERS) technique, for the highly sensitive biological analysis in a microfluidic sensor, has been developed in our research group. Here, a highly precise quantitative measurement can be obtained if continuous flow and homogeneous mixing condition between analytes and silver nano-colloids are maintained. Recently, we also reported a new analytical method of DNA hybridization involving a PDMS microfluidic sensor using fluorescence energy transfer (FRET). This method overcomes many of the drawbacks of microarray chips, such as long hybridization times and inconvenient immobilization procedures. In this paper, our recent applications of the confocal Raman/fluorescence microscopic technology to a highly sensitive lab-on-a-chip detection will be reviewed.

Epidemiologic Study of Human Epidermal Growth Factor Receptor 2 Expression in Advanced/Metastatic Gastric Cancer: an Assessment of Human Epidermal Growth Factor Receptor 2 Status in Tumor Tissue Samples of Gastric and Gastro-Esophageal Junction Cancer

  • Seo, Kyung Won;Jeon, Taeyong;Kim, Sewon;Kim, Sung Soo;Kim, Kwanghee;Suh, Byoung-Jo;Hwang, Sunhwi;Choi, SeongHee;Ryu, Seungwan;Min, Jae Seok;Lee, Young-Joon;Jee, Ye Seob;Chae, Hyeondong;Yang, Doo Hyun;Lee, Sang Ho
    • Journal of Gastric Cancer
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    • v.17 no.1
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    • pp.52-62
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    • 2017
  • Purpose: The Trastuzumab for gastric cancer (GC) trial identified human epidermal growth factor receptor 2 (HER2) as a predictor of successful treatment with trastuzumab (HER2 receptor targeting agent) among patients with advanced/metastatic GC. To date, the prevalence of HER2 overexpression in the Korean population is unknown. The present study aimed to assess the incidence of HER2 positivity among GC and gastroesophageal (GE) junction cancer samples and the relationship between HER2 overexpression and clinicopathological characteristics in Korean patients. Materials and Methods: Tumor samples collected from 1,695 patients with histologically proven GC or GE junction enrolled at 14 different hospitals in Korea were examined. After gathering clinicopathological data of all patients, HER2 status was assessed by immunohistochemistry (IHC) at each hospital, and IHC 2+ cases were subjected to silver-enhanced in situ hybridization at 3 central laboratories. Results: A total of 182 specimens tested positive for HER2, whereas 1,505 tested negative. Therefore, the overall HER2-positive rate in this study was 10.8% (95% confidence interval=9.3%-12.3%). The HER2-positive rate was higher among intestinal-type cases (17.6%) than among other types, and was higher among patients older than 70 years and 50 years of age, compared to other age groups. Conclusions: Our evaluation of the HER2 positivity rate (10.8%) among Korean patients with GC and GE junction indicated the necessity of epidemiological data when conducting studies related to HER2 expression in GC and GE junction.

Ligand-Independent Epidermal Growth Factor Receptor Overexpression Correlates with Poor Prognosis in Colorectal Cancer

  • Yun, Sumi;Kwak, Yoonjin;Nam, Soo Kyung;Seo, An Na;Oh, Heung-Kwon;Kim, Duck-Woo;Kang, Sung-Bum;Lee, Hye Seung
    • Cancer Research and Treatment
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    • v.50 no.4
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    • pp.1351-1361
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    • 2018
  • Purpose Molecular treatments targeting epidermal growth factor receptors (EGFRs) are important strategies for advanced colorectal cancer (CRC). However, clinicopathologic implications of EGFRs and EGFR ligand signaling have not been fully evaluated. We evaluated the expression of EGFR ligands and correlation with their receptors, clinicopathologic factors, and patients' survival with CRC. Materials and Methods The expression of EGFR ligands, including heparin binding epidermal growth factor-like growth factor (HBEGF), transforming growth factor (TGF), betacellulin, and epidermal growth factor (EGF), were evaluated in 331 consecutive CRC samples using mRNA in situ hybridization (ISH). We also evaluated the expression status of EGFR, human epidermal growth factor receptor 2 (HER2), HER3, and HER4 using immunohistochemistry and/or silver ISH. Results Unlike low incidences of TGF (38.1%), betacellulin (7.9%), and EGF (2.1%), HBEGF expression was noted in 62.2% of CRC samples. However, the expression of each EGFR ligand did not reveal significant correlations with survival. The combined analyses of EGFR ligands and EGFR expression indicated that the ligands-/EGFR+ group showed a significant association with the worst disease-free survival (DFS; p=0.018) and overall survival (OS; p=0.005). It was also an independent, unfavorable prognostic factor for DFS (p=0.026) and OS (p=0.007). Additionally, HER4 nuclear expression, regardless of ligand expression, was an independent, favorable prognostic factor for DFS (p=0.034) and OS (p=0.049), by multivariate analysis. Conclusion Ligand-independent EGFR overexpression was suggested to have a significant prognostic impact; thus, the expression status of EGFR ligands, in addition to EGFR, might be necessary for predicting patients' outcome in CRC.

Is Helicobacter pylori a Poor Prognostic Factor for HER-2 SISH Positive Gastric Cancer?

  • Selcukbiricik, Fatih;Tural, Deniz;Erdamar, Sibel;Buyukunal, Evin;Demirelli, Fuat;Serdengecti, Suheyla
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.5
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    • pp.3319-3322
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    • 2013
  • Background: Helicobacter pylori (H. pylori) is one of the risk factors for gastric cancer (GC). Any prognostic effect of HER-2 status in gastric lymph node metastasis in H. pylori positive cases is unknown. Materials and Methods: A total of 74 patients, 47 (64%) male, and 27 (34%) female, who had subtotal or total gastrectomy and also positive lymph nodes, were included in the study. Age range was 29-87 years, and median age was 58 years. HER-2 expression was assessed in both gastric resection samples and lymph node material with carcinoma metastasis of the same patient by immunohistochemistry (IHC) and silver in situ hybridization (SISH) methods. H. pylori status was examined in gastric materials of all patients. Relationships between HER-2 status in gastric cancers and lymph nodes and H. pylori status were investigated. Results: H. pylori was positive in 40 cases (54%), and negative in 34 (46%). While in the primary tissues of H. pylori positive cases, SISH positivity for HER-2 was observed in 13 cases (86%), SISH negativity was observed in 2 (14%), in metastatic lymph nodes 21 cases (72%) were SISH positive and 8 cases (28%) were SISH negative (P=0.005 and P=0.019, respectively). Initial CEA values were high in 18 cases (78%) with positive H. pylori and in 5 cases (22%) with negative H. pylori (P=0.009). While SISH data of patients were negative in 59 cases (80%) and positive in 15 cases (20%) in primary tissues, they were negative in 56 cases (75%) and positive in 18 cases (25%) in lymph nodes. Discrepancy between primary tissue and lymph node results was detected in 3 cases, in which SISH was negative in the primary tissue and HER-2 expression was positive in the lymph nodes. Conclusions: Clinical progression was poor in H. pylori positive cases with HER-2 negativity in primary gastric tissue, but HER-2 positivity in the lymph nodes. SISH positivity can be expected in H. pylori positive cases, and it may be predicted that these cases can benefit from trastuzumab treatment.

Factors that Predict Clinical Benefit of EGFR TKI Therapy in Patients with EGFR Wild-Type Lung Adenocarcinoma

  • Kim, Seo Yun;Myung, Jae Kyung;Kim, Hye-Ryoun;Na, Im Il;Koh, Jae Soo;Baek, Hee Jong;Kim, Cheol Hyeon
    • Tuberculosis and Respiratory Diseases
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    • v.82 no.1
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    • pp.62-70
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    • 2019
  • Background: Epidermal growth factor receptor (EGFR) mutations in non-small cell lung cancers have emerged as key predictive biomarkers in EGFR tyrosine kinase inhibitor (TKI) treatment. However, a few patients with wild-type EGFR also respond to EGFR TKIs. This study investigated the factors predicting successful EGFR TKI treatment in lung adenocarcinoma patients with wild-type EGFR. Methods: We examined 66 patients diagnosed with lung adenocarcinoma carrying wide-type EGFR who were treated with EGFR TKIs. The EGFR gene copy number was assessed by silver in situ hybridization (SISH). We evaluated the clinical factors and EGFR gene copy numbers that are associated with a favorable clinical response to EGFR TKIs. Results: The objective response rate was 12.1%, while the disease control rate was 40.9%. EGFR SISH analysis was feasible in 23 cases. Twelve patients tested EGFR SISH-positive, and 11 were EGFR SISH-negative, with no significant difference in tumor response and survival between EGFR SISH-positive and -negative patients. The overall median progression-free survival (PFS) and overall survival (OS) of 66 patients were 2.1 months and 9.7 months, respectively. Female sex and Eastern Cooperative Oncology Group (ECOG) performance status (PS) of 0-1 were independent predictors of PFS. ECOG PS 0-1 and a low tumor burden of extrathoracic metastasis were independent predictors of good OS. Conclusion: Factors such as good PS, female sex, and low tumor burden may predict favorable outcomes following EGFR TKI therapy in patients with EGFR wild-type lung adenocarcinoma. However, EGFR gene copy number was not predictive of survival.