<i>KRAS</i>fluorescence in situ hybridisation testing for the detection and diagnosis of pancreatic adenocarcinoma

Shiroma, Noriyuki; Arihiro, Koji; Oda, Miyo; Orita, Makoto

doi:10.1136/jclinpath-2018-205002

Cited by 3 publications

(3 citation statements)

References 34 publications

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“…Wide regions of DNA (kilobases to megabases) containing these oncogenes can be organized as extrachromosomal copies (double minutes), as tandem repeats within a chromosome or may be distributed at various sites within the genome [78]. KRAS amplifications has been reported in various malignancies, primarily in pancreatic [59, 79], colorectal [80, 81] and lung [58, 82] tumors, but also have been noted in gastric [83], ovarian [84] and endometrial [85] malignancies, but less frequently in prostate cancers [46]. In our study, the data did not suggest either amplification of large chromosomal portions of DNA encapsulating the KRAS gene, or chromosome duplication (e.g.…”

Section: Discussionmentioning

confidence: 99%

Arsenite malignantly transforms human prostate epithelial cells in vitro by gene amplification of mutated KRAS

et al. 2019

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Inorganic arsenic is an environmental human carcinogen of several organs including the urinary tract. RWPE-1 cells are immortalized, non-tumorigenic, human prostate epithelia that become malignantly transformed into the CAsE-PE line after continuous in vitro exposure to 5μM arsenite over a period of months. For insight into in vitro arsenite transformation, we performed RNA-seq for differential gene expression and targeted sequencing of KRAS. We report >7,000 differentially expressed transcripts in CAsE-PE cells compared to RWPE-1 cells at >2-fold change, q<0.05 by RNA-seq. Notably, KRAS expression was highly elevated in CAsE-PE cells, with pathway analysis supporting increased cell proliferation, cell motility, survival and cancer pathways. Targeted DNA sequencing of KRAS revealed a mutant specific allelic imbalance, ‘MASI’, frequently found in primary clinical tumors. We found high expression of a mutated KRAS transcript carrying oncogenic mutations at codons 12 and 59 and many silent mutations, accompanied by lower expression of a wild-type allele. Parallel cultures of RWPE-1 cells retained a wild-type KRAS genotype. Copy number analysis and sequencing showed amplification of the mutant KRAS allele. KRAS is expressed as two splice variants, KRAS4a and KRAS4b, where variant 4b is more prevalent in normal cells compared to greater levels of variant 4a seen in tumor cells. 454 Roche sequencing measured KRAS variants in each cell type. We found KRAS4a as the predominant transcript variant in CAsE-PE cells compared to KRAS4b, the variant expressed primarily in RWPE-1 cells and in normal prostate, early passage, primary epithelial cells. Overall, gene expression data were consistent with KRAS-driven proliferation pathways found in spontaneous tumors and malignantly transformed cell lines. Arsenite is recognized as an important environmental carcinogen, but it is not a direct mutagen. Further investigations into this in vitro transformation model will focus on genomic events that cause arsenite-mediated mutation and overexpression of KRAS in CAsE-PE cells.

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Section: Discussionmentioning

confidence: 99%

Arsenite malignantly transforms human prostate epithelial cells in vitro by gene amplification of mutated KRAS

et al. 2019

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“…KRAS mutations have been well characterized in the pathogenesis of pancreatic cancer [158,159]. Alterations in KRAS are thought to be an early event in IPMN biogenesis as it is found in all types without significant differences [160,161].…”

Section: Genomic Markersmentioning

confidence: 99%

Elevating pancreatic cystic lesion stratification: Current and future pancreatic cancer biomarker(s)

Carmicheal

Patel

Dalal

et al. 2020

Biochimica et Biophysica Acta (BBA) - Reviews on Cancer

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Pancreatic ductal adenocarcinoma (PDAC) is an incredibly deadly disease with a 5-year survival rate of 9%. The presence of pancreatic cystic lesions (PCLs) confers an increased likelihood of future pancreatic cancer in patients placing them in a high-risk category. Discerning concurrent malignancy and risk of future PCL progression to cancer must be carefully and accurately determined to improve survival outcomes and avoid unnecessary morbidity of pancreatic resection. Unfortunately, current image-based guidelines are inadequate to distinguish benign from malignant lesions. There continues to be a need for accurate molecular and imaging biomarker(s) capable of identifying malignant PCLs and predicting the malignant potential of PCLs to enable risk stratification and effective intervention management. This review provides an update on the current status of biomarkers from pancreatic cystic fluid, pancreatic juice, and seromic molecular *

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“…A fundamental benefit of FISH is that several probes can be multiplexed to concurrently assess multiple genes, regions, or chromosomes [2,38,87]. For example, Shiroma and colleagues multiplexed KRAS and centromere (chromosome 12) probes to correlate gene amplification (S-CIN) and whole chromosome copy number changes (N-CIN) with tumor stage, grade, and survival in pancreatic ductal adenocarcinomas [93], while Penner-Goeke et al multiplexed three distinct CEPs to determine the prevalence and dynamics of N-CIN within recurrent and drug-resistant high-grade serous ovarian cancer [94]. Unfortunately, multiplexing is typically limited to ≤ 4 probes due to the limited number of distinct fluorophores that can be individually imaged without spectral overlap on most conventional fluorescent microscopes [94,95].…”

Section: Cytogenetic Approachesmentioning

confidence: 99%

Detecting Chromosome Instability in Cancer: Approaches to Resolve Cell-to-Cell Heterogeneity

Lepage

Morden

Palmer

et al. 2019

Cancers

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Chromosome instability (CIN) is defined as an increased rate of chromosome gains and losses that manifests as cell-to-cell karyotypic heterogeneity and drives cancer initiation and evolution. Current research efforts are aimed at identifying the etiological origins of CIN, establishing its roles in cancer pathogenesis, understanding its implications for patient prognosis, and developing novel therapeutics that are capable of exploiting CIN. Thus, the ability to accurately identify and evaluate CIN is critical within both research and clinical settings. Here, we provide an overview of quantitative single cell approaches that evaluate and resolve cell-to-cell heterogeneity and CIN, and discuss considerations when selecting the most appropriate approach to suit both research and clinical contexts.

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KRASfluorescence in situ hybridisation testing for the detection and diagnosis of pancreatic adenocarcinoma

Cited by 3 publications

References 34 publications

Arsenite malignantly transforms human prostate epithelial cells in vitro by gene amplification of mutated KRAS

Arsenite malignantly transforms human prostate epithelial cells in vitro by gene amplification of mutated KRAS

Elevating pancreatic cystic lesion stratification: Current and future pancreatic cancer biomarker(s)

Detecting Chromosome Instability in Cancer: Approaches to Resolve Cell-to-Cell Heterogeneity

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