Cytologic evidence for gene amplification in methotrexate-resistant cells obtained from a patient with ovarian adenocarcinoma.

1984 ◽  
Vol 2 (1) ◽  
pp. 8-15 ◽  
Author(s):  
J M Trent ◽  
R N Buick ◽  
S Olson ◽  
R C Horns ◽  
R T Schimke
Keyword(s):  
Gene Amplification ◽  
Gene Dosage ◽  
Dhfr Gene ◽  
Ovarian Adenocarcinoma ◽  
Serous Adenocarcinoma ◽  
Ovarian Serous Adenocarcinoma ◽  
Colony Forming Units ◽  
Well Differentiated ◽  
Free Media

To analyze if methotrexate (MTX) resistance arises from gene amplification in a patient treated clinically with MTX, the cytogenetic and drug sensitivity profile of the tumor colony forming units (TCFUs) from a 58-year-old woman with stage III well-differentiated ovarian serous adenocarcinoma was studied. This patient had not received treatment directed against her tumor for nine months before this study, but had received oral-dose MTX (2.5 mg, twice weekly) for three years for the treatment of psoriasis. Analysis of TCFUs grown in nucleoside-free media demonstrated MTX resistance at concentrations of up to 100 micrograms/mL (2.2 X 10(-4)M). Cytologic evidence for dihydrofolate reductase (DHFR) gene amplification in TCFUs was determined by in situ hybridization, using radiolabeled cDNA to DHFR mRNA. Results localized the DHFR sequences to an abnormally staining region present on chromosome 4q. This study supports the notion that alterations in gene dosage (that is, gene amplification) play a role in the development of drug resistance in spontaneous human cancers.

scholarly journals Dedifferentiated Liposarcoma Arising in an Esophageal Polyp: A Case Report

2012 ◽  
Vol 2012 ◽  
pp. 1-3 ◽  
Author(s):  
Jorge Torres-Mora ◽  
Ann Moyer ◽  
Mark Topazian ◽  
Jeffrey Alexander ◽  
Tsung-Teh Wu ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Case Report ◽  
Fluorescence In Situ Hybridization ◽  
Gene Amplification ◽  
Dedifferentiated Liposarcoma ◽  
Well Differentiated

Liposarcoma is one of the most common sarcomas in adults, but only rarely presents as an esophageal primary. There have been several reports of well-differentiated liposarcoma (WDL) arising in the esophagus, but we present a case of dedifferentiated liposarcoma (DL) presenting as a large esophageal polyp. We believe this is the first reported case of DL of the esophagus with morphologic evidence of both well-differentiated and dedifferentiated components. The diagnosis was confirmed by demonstration of CPM gene amplification by fluorescence in situ hybridization (FISH).

scholarly journals FluorescenceIn SituHybridization forMDM2Amplification as a Routine Ancillary Diagnostic Tool for Suspected Well-Differentiated and Dedifferentiated Liposarcomas: Experience at a Tertiary Center

Sarcoma ◽  
2015 ◽  
Vol 2015 ◽  
pp. 1-10 ◽  
Author(s):  
Khin Thway ◽  
Jayson Wang ◽  
John Swansbury ◽  
Toon Min ◽  
Cyril Fisher
Keyword(s):  
Gene Amplification ◽  
Diagnostic Tool ◽  
Spindle Cell ◽  
Significant Group ◽  
Lipomatous Tumor ◽  
Tertiary Center ◽  
Benign Histology ◽  
Well Differentiated ◽  
Correlation Rate

Background. The assessment ofMDM2gene amplification by fluorescencein situhybridization (FISH) has become a routine ancillary tool for diagnosing atypical lipomatous tumor (ALT)/well-differentiated liposarcoma and dedifferentiated liposarcoma (WDL/DDL) in specialist sarcoma units. We describe our experience of its utility at our tertiary institute.Methods. All routine histology samples in whichMDM2amplification was assessed with FISH over a 2-year period were included, and FISH results were correlated with clinical and histologic findings.Results. 365 samples from 347 patients had FISH forMDM2gene amplification. 170 were positive (i.e., showedMDM2gene amplification), 192 were negative, and 3 were technically unsatisfactory. There were 122 histologically benign cases showing a histology:FISH concordance rate of 92.6%, 142 WDL/DDL (concordance 96.5%), and 34 cases histologically equivocal for WDL (concordance 50%). Of 64 spindle cell/pleomorphic neoplasms (in which DDL was a differential diagnosis), 21.9% showedMDM2amplification. Of the cases with discrepant histology and FISH, all but 3 had diagnoses amended following FISH results. For discrepancies of benign histology but positive FISH, lesions were on average larger, more frequently in “classical” (intra-abdominal or inguinal) sites for WDL/DDL and more frequently core biopsies. Discrepancies of malignant histology but negative FISH were smaller, less frequently in “classical” sites but again more frequently core biopsies.Conclusions. FISH has a high correlation rate with histology for cases with firm histologic diagnoses of lipoma or WDL/DDL. It is a useful ancillary diagnostic tool in histologically equivocal cases, particularly in WDL lacking significant histologic atypia or DDL without corresponding WDL component, especially in larger tumors, those from intra-abdominal or inguinal sites or core biopsies. There is a significant group of well-differentiated adipocytic neoplasms which are difficult to diagnose on morphology alone, in which FISH forMDM2amplification is diagnostically contributory.

The genes CNV features in the tumor cells in patients with ovarian serous adenocarcinoma.

2020 ◽  
Vol 38 (15_suppl) ◽  
pp. e13641-e13641
Author(s):  
Marieta R. Tsandekova ◽  
Natalya V. Porkhanova ◽  
Oleg Ivanovich Kit ◽  
Denis S. Kutilin
Keyword(s):  
Cluster Analysis ◽  
Epithelial Cells ◽  
Early Diagnosis ◽  
Tumor Cells ◽  
Copy Number ◽  
Molecular Mechanisms ◽  
Ovarian Adenocarcinoma ◽  
Serous Adenocarcinoma ◽  
Ovarian Serous Adenocarcinoma ◽  
Ovarian Epithelial Cells

e13641 Background: Serous adenocarcinoma is the most common subtype of ovarian cancer. Early diagnosis of this disease can significantly improve the prognosis. Development of more effective methods for early diagnosis and identification of more reliable markers require understanding the molecular mechanisms underlying the malignant behavior of ovarian epithelial cells. The aim of the study was to analyze the copy number variation (CNV) of genes that regulate apoptosis, DNA repair, cell proliferation, metabolism and estrogen reception in tumor and normal ovarian cells. Methods: For the study, we used tissue sections from FFPE blocks of 65 patients with serous ovarian adenocarcinoma diagnosis. Tumor and normal ovarian epithelial cells were isolated using laser microdissection with non-contact capture (Palm MicroBeam, Carl Zeiss). DNA was extracted from cells using the phenol-chloroform method. CNV of 33 genes (BRAF, KRAS, EGFR, PIK3CA, PTEN, TP53, BRCA1, BRCA2, PRKCI, NOTCH1, AKT1, BAX, CASP7, CASP3, CASP8, CASP9, MDM2, BCL2, CYP1A1, CYP1A1, CYP1A2, CYP1A2, CYP1A2, CYP1A1, CYP1A2, CYP1, ESR2, GPER, STS, SULT1A, SULT1E1, OCT4, SOX2, C-MYC, SOX18, SCNN1A) were determined by Real-Time qPCR method (reference B2M, GAPDH). Statistical analysis was performed using the Mann-Whitney test. For cluster analysis (Hierarchical Clustering, Euclidean distance) scripts in R were used. Results: A statistically significant (p < 0.005) increase in the CNV of PTEN, MDM2, SOX2, CYP1B1, ESR1, and SULT1E1 by 2.0, 2.0, 1.8, 2.5, 3.0 and 2.0 times, respectively, was found, as well as a 2.0-fold decrease in the CNV of CASP3 and CASP8 in tumor cells relatively to normal ones. Cluster analysis allowed us to distinguish 2 groups of serous adenocarcinoma samples that differed in gene CNV (p < 0.005): in group 1 (n = 40), the copy number of the SOX2, MDM2, ESR1, CYP1B1 and SULT1E1 genes was increased and the copy number of the TP53 and BRCA2 genes was reduced, in group 2 (n = 25), copy number of PTEN, PIK3CA, BCL2 was increased and copy number of BAX, CASP3 and CASP8 was lower. Conclusions: CNV analysis revealed the most characteristic markers of ovarian serous adenocarcinoma cells. Based on the differential CNV, 2 molecular subtypes of serous adenocarcinoma were distinguished.

Evaluation of Stability in the DHFR Gene Amplification System Using Fluorescence In Situ Hybridization

2006 ◽  
pp. 259-263
Author(s):  
Fumi Nakanishi ◽  
Tomohiro Yoshikawa ◽  
Seima Itami ◽  
Takeshi Omasa ◽  
Yoshio Katakura ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Gene Amplification ◽  
Dhfr Gene ◽  
Amplification System

A PILOT STUDY OF HER-2/NEU GENE AMPLIFICATION ASSESSED BY FLUORESCENCE IN SITU HYBRIDIZATION (FISH) IN GASTRIC CANCER

2014 ◽  
pp. 15-20
Author(s):  
Van Huy Tran ◽  
Thi Minh Thi Ha ◽  
Trung Nghia Van ◽  
Viet Nhan Nguyen ◽  
Phan Tuong Quynh Le ◽  
...  
Keyword(s):  
Gastric Cancer ◽  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Cancer Patients ◽  
Gene Amplification ◽  
Cancer Tissue ◽  
Tissue Samples ◽  
Her 2 ◽  
Gastric Cancer Patients

Background: HER-2/neu is a predictive biomarker for treatment of gastric cancer using trastuzumab in combination with chemotherapy. This study aimed to evaluate the status of HER-2/neu gene amplification using fluorescence in situ hybridization (FISH) in gastric cancer. Patients and methods: thirty six gastric cancer patients were assessed HER-2/neu gene amplification by FISH using PathVysionTM HER-2 DNA Probe kit (including HER-2/neu probe and CEP-17 probe) with biopsy and surgical specimens. Results: The HER-2/neu gene amplification was observed in three cases (8.3%), the HER-2/neu gene amplification rate in Lauren’s intestinal-type and diffuse-type were 11.8% and 5.2%, respectively. Conclusion: We applied successfully FISH technique with gastric cancer tissue samples. This technique could be performed as routine test in gastric cancer in order to select patients that benefit from trastuzumab in combination with chemotherapy.

THE ASSOCIATION OF THE APOLIPOPROTEIN E4 ALLELE (RS 429358) WITH OVARIAN SEROUS ADENOCARCINOMA

2017 ◽  
Vol 63 (4) ◽  
pp. 627-631
Author(s):  
Tatyana Ivanova ◽  
Nataliya Sychenkova ◽  
Vera Khorokhorina ◽  
Nikolay Ryabchenko ◽  
Sergey Ivanov ◽  
...  
Keyword(s):  
Older Women ◽  
Odds Ratio ◽  
Area Under Curve ◽  
Reproductive Age ◽  
Prognostic Indicators ◽  
Separate Analysis ◽  
Women Of Reproductive Age ◽  
Serous Adenocarcinoma ◽  
Ovarian Serous Adenocarcinoma ◽  
E4 Allele

The distribution of APOE 4 allele (rs 429358, C) was analyzed in healthy women (N=454) and patients with ovarian serous adenocarcinoma (N=114) in order to identify genetic predisposition to the disease. We determined the prognostic indicators of the E4 allele as a marker: odds ratio (OR) and AUC (Area Under Curve) - an area under the ROC curve. It was shown that APOE 4 allele was significantly associated with ovarian serous adenocarcinoma (p = 0,003; 0R=1,94; AUC=0,55). The Е4 genotypes frequency was significantly increased among patients (p = 0.02; 0R=1,8). Separate analysis of the two age subgroups (over 46 years and younger) found that the chance of developing ovarian serous adenocarcinoma was significantly increased for older women (p = 0,006; OR = 2,24, AUC = 0,76). Possible associations of APOE 4 with the ovarian serous adenocarcinoma in women of reproductive age deserve further studying.

Laboratory Testing for HER2/neu in Breast Carcinoma: An Evolving Strategy to Predict Response to Targeted Therapy

2001 ◽  
Vol 8 (5) ◽  
pp. 415-418 ◽  
Author(s):  
Nils M. Diaz
Keyword(s):  
Targeted Therapy ◽  
Breast Carcinoma ◽  
Gene Amplification ◽  
Laboratory Testing ◽  
False Positives ◽  
Response To Therapy ◽  
Fish Analysis ◽  
Breast Carcinomas ◽  
Testing Strategies

Background Laboratory testing of HER2/neu in breast carcinoma has become vital to patient care following the approval of trastuzumab as the first therapy to target the HER2/neu oncoprotein. Initial clinical trials used immunohistochemistry (IHC) to test for HER2/neu overexpression in order to select patients for therapy. Fluorescence in situ hybridization (FISH), which tests for gene amplification, is more specific and sensitive than IHC when either assay is compared with HER2/neu overexpression as determined by Northern or Western blot analysis. Many weak overexpressors on IHC testing are not gene amplified on FISH analysis. Such weak overexpressors may be considered false-positives and raise the question of how best to test for HER2/neu. Methods The literature was surveyed regarding testing for HER2/neu overexpression in breast carcinomas and alternative testing strategies. Results False-positive results are a significant problem when IHC is exclusively used to test for HER2/neu overexpression. The false-positives are overwhelmingly confined to the group of 2+ positives and do not respond to targeted therapy. In contrast, concordance between IHC and FISH is high when immunostaining is interpreted as either negative or strongly positive (3+). Whereas some recent studies have suggested that FISH may better predict response to anti-HER2/neu therapy than IHC, others have indicated that IHC is as effective a predictor as FISH. IHC is less technically demanding and costly than FISH. Conclusions IHC analysis of HER2/neu in breast carcinoma is a useful predictor of response to therapy with trastuzumab when strongly positive. Negative immunostaining is highly concordant with a lack of gene amplification by FISH. Most weakly positive overexpressors are false-positives on testing with FISH. Thus, screening of breast carcinomas with IHC and confirmation of weakly positive IHC results by FISH is an effective evolving strategy for testing HER2/neu as a predictor of response to targeted therapy.

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