Dataset Information

Detection of CCNE1/URI (19q12) amplification by in situ hybridisation is common in high grade and type II endometrial cancer.

ABSTRACT: One TCGA subgroup of endometrial cancer (EC) is characterised by extensive genomic DNA copy number alterations. CCNE1 located at 19q12 is frequently amplified in EC and a target for anti-cancer therapy. The relevance of URI, also located at 19q12, is unknown. To evaluate the prevalence of 19q12 (CCNE1/URI) in EC, we investigated different histologic types by in situ hybridisation (ISH) and copy number assay. We applied a previously established 19q12 ISH for the detection of CCNE1/URI copy numbers in EC (n = 270) using conventional bright field microscopy. In a subset (n = 21), 19q12 amplification status was validated by OncoScan assay. Manual ISH was controlled by a recently developed computational ISHProfiler algorithm. Associations of 19q12 status with Cyclin E1, URI and p53 expression, and clinico-pathological parameters were tested.Amplification of 19q12 (CCNE1/URI) was found in 10.4% (28/270) and was significantly associated with type II EC (high grade and non-endometrioid; p < 0.0001), advanced FIGO stage (p = 0.001), high Cyclin E1 expression (p = 0.008) and aberrant p53 expression (p = 0.04). 19q12 ISH data were confirmed by OncoScan and computational ISHProfiler techniques. The 19q12 in situ hybridisation is a feasible and robust biomarker assay in molecular pathology. Amplification of CCNE1/URI predominantly occurred in type II endometrial cancer. Prospective clinical trials are warranted to assess the utility of combined 19q12 amplification and Cyclin E1/URI protein expression analysis for the prediction of therapeutic response to chemotherapy and/or cyclin-dependent kinase inhibitors in patients with endometrial cancer.

SUBMITTER: Noske A

PROVIDER: S-EPMC5362444 | biostudies-literature | 2017 Feb

REPOSITORIES: biostudies-literature

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Detection of CCNE1/URI (19q12) amplification by in situ hybridisation is common in high grade and type II endometrial cancer.

Noske Aurelia A Brandt Simone S Valtcheva Nadejda N Wagner Ulrich U Zhong Qing Q Bellini Elisa E Fink Daniel D Obermann Ellen C EC Moch Holger H Wild Peter J PJ

Oncotarget 20170201 9

One TCGA subgroup of endometrial cancer (EC) is characterised by extensive genomic DNA copy number alterations. CCNE1 located at 19q12 is frequently amplified in EC and a target for anti-cancer therapy. The relevance of URI, also located at 19q12, is unknown. To evaluate the prevalence of 19q12 (CCNE1/URI) in EC, we investigated different histologic types by in situ hybridisation (ISH) and copy number assay. We applied a previously established 19q12 ISH for the detection of CCNE1/URI copy number ...[more]

PMID: 27582547

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Project description:In this study, we analysed gene amplification, RNA expression and protein expression of the c-myc gene on archival tissue specimens of high-grade human breast cancer, using fluorescent in situ hybridisation (FISH), nonradioactive in situ hybridisation and immunohistochemistry. The specific question that we addressed was whether expression of c-Myc mRNA and protein were correlated with its gene copy amplification, as determined by FISH. Although c-Myc is one of the most commonly amplified oncogenes in human breast cancer, few studies have utilised in situ approaches to directly analyse the gene copy amplification, RNA transcription and protein expression on human breast tumour tissue sections. We now report that by using the sensitive FISH technique, a high proportion (70%) of high-grade breast carcinoma were amplified for the c-myc gene, irrespective of status of the oestrogen receptor. However, the level of amplification was low, ranging between one and four copies of gene gains, and the majority (84%) of the cases with this gene amplification gained only one to two copies. Approximately 92% of the cases were positive for c-myc RNA transcription, and essentially all demonstrated c-myc protein expression. In fact, a wide range of expression levels were detected. Statistically significant correlations were identified among the gene amplification indices, the RNA expression scores and protein expression scores. c-myc gene amplification, as detected by FISH, was significantly associated with expression of its mRNA, as measured by the intensity of in situ hybridisation in invasive cells (P=0.0067), and by the percentage of invasive cells positive for mRNA expression (P=0.0006). c-myc gene amplification was also correlated with the percentage of tumour cells which expressed high levels of its protein, as detected by immunohistochemistry in invasive cells (P=0.0016). Thus, although multiple mechanisms are known to regulate normal and aberrent expression of c-myc, in this study, where in situ methodologies were used to evaluate high-grade human breast cancers, gene amplification of c-myc appears to play a key role in regulating expression of its mRNA and protein.

Project description:INTRODUCTION:The 19q12 locus is amplified in a subgroup of oestrogen receptor (ER)-negative grade III breast cancers. This amplicon comprises nine genes, including cyclin E1 (CCNE1), which has been proposed as its 'driver'. The aim of this study was to identify the genes within the 19q12 amplicon whose expression is required for the survival of cancer cells harbouring their amplification. METHODS:We investigated the presence of 19q12 amplification in a series of 313 frozen primary breast cancers and 56 breast cancer cell lines using microarray comparative genomic hybridisation (aCGH). The nine genes mapping to the smallest region of amplification on 19q12 were silenced using RNA interference in phenotypically matched breast cancer cell lines with (MDA-MB-157 and HCC1569) and without (Hs578T, MCF7, MDA-MB-231, ZR75.1, JIMT1 and BT474) amplification of this locus. Genes whose silencing was selectively lethal in amplified cells were taken forward for further validation. The effects of cyclin-dependent kinase 2 (CDK2) silencing and chemical inhibition were tested in cancer cells with and without CCNE1 amplification. RESULTS:19q12 amplification was identified in 7.8% of ER-negative grade III breast cancer. Of the nine genes mapping to this amplicon, UQCRFS1, POP4, PLEKHF1, C19ORF12, CCNE1 and C19ORF2 were significantly over-expressed when amplified in primary breast cancers and/or breast cancer cell lines. Silencing of POP4, PLEKHF1, CCNE1 and TSZH3 selectively reduced cell viability in cancer cells harbouring their amplification. Cancer cells with CCNE1 amplification were shown to be dependent on CDK2 expression and kinase activity for their survival. CONCLUSIONS:The 19q12 amplicon may harbour more than a single 'driver', given that expression of POP4, PLEKHF1, CCNE1 and TSZH3 is required for the survival of cancer cells displaying their amplification. The observation that cancer cells harbouring CCNE1 gene amplification are sensitive to CDK2 inhibitors provides a rationale for the testing of these chemical inhibitors in a subgroup of patients with ER-negative grade III breast cancers.

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Dataset Information

Detection of CCNE1/URI (19q12) amplification by in situ hybridisation is common in high grade and type II endometrial cancer.

Publications

Detection of CCNE1/URI (19q12) amplification by in situ hybridisation is common in high grade and type II endometrial cancer.

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