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      RAS Mutational Status Detection in Tissue, Plasma, and Stool Samples for Colorectal Cancer

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          Abstract

          Objective

          RAS gene testing on tumor tissue biopsies is required for metastatic colorectal cancer (CRC) patients. However, it is infeasible for patients after curative surgery and repeated biopsy. This study is aimed at evaluating the consistency of RAS genes in patient's plasma, stool, and tumor tissue samples, to explore whether plasma and stool samples can supplement or replace tumor tissue to assess baseline RAS gene status.

          Methods

          Between June 2016 and October 2017, 53 patients with stage I-IV CRC from the Liaoning Cancer Hospital and the Department of Medical Oncology of the First Hospital of China Medical University were enrolled in the study. Patient tissues, peripheral blood, and stool samples were collected, and RAS gene tests were performed.

          Results

          Analysis of the KRAS gene in tissue, plasma, and stool samples from 53 CRC patients detected 25 cases (47%) of KRAS gene mutations in the tissue samples, 20 cases (38%) of KRAS gene mutations in plasma, and 18 (34%) KRAS gene mutations in fecal samples. The overall consistency of KRAS gene status between tissue samples and plasma samples was 77.4% ( p ≤ 0.05) and between tissue samples and stool samples was 83% ( p ≤ 0.05). In stage IV cases, the agreement of KRAS gene status between tissue and plasma samples was 93.8% ( p ≤ 0.05) and 93.8% ( p ≤ 0.05) between tissue and stool samples.

          Conclusion

          There was a high overall consistency in KRAS mutational assessment between plasma, stool, and tissue samples. In stage IV patients, the consistency of KRAS gene detection between tissue and stools or plasma was higher.

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          Most cited references13

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          Plasma ctDNA RAS mutation analysis for the diagnosis and treatment monitoring of metastatic colorectal cancer patients

          Abstract Background RAS assessment is mandatory for therapy decision in metastatic colorectal cancer (mCRC) patients. This determination is based on tumor tissue, however, genotyping of circulating tumor (ct)DNA offers clear advantages as a minimally invasive method that represents tumor heterogeneity. Our study aims to evaluate the use of ctDNA as an alternative for determining baseline RAS status and subsequent monitoring of RAS mutations during therapy as a component of routine clinical practice. Patients and methods RAS mutational status in plasma was evaluated in mCRC patients by OncoBEAM™ RAS CRC assay. Concordance of results in plasma and tissue was retrospectively evaluated. RAS mutations were also prospectively monitored in longitudinal plasma samples from selected patients. Results Analysis of RAS in tissue and plasma samples from 115 mCRC patients showed a 93% overall agreement. Plasma/tissue RAS discrepancies were mainly explained by spatial and temporal tumor heterogeneity. Analysis of clinico-pathological features showed that the site of metastasis (i.e. peritoneal, lung), the histology of the tumor (i.e. mucinous) and administration of treatment previous to blood collection negatively impacted the detection of RAS in ctDNA. In patients with baseline mutant RAS tumors treated with chemotherapy/antiangiogenic, longitudinal analysis of RAS ctDNA mirrored response to treatment, being an early predictor of response. In patients RAS wt, longitudinal monitoring of RAS ctDNA revealed that OncoBEAM was useful to detect emergence of RAS mutations during anti-EGFR treatment. Conclusion The high overall agreement in RAS mutational assessment between plasma and tissue supports blood-based testing with OncoBEAM™ as a viable alternative for genotyping RAS of mCRC patients in routine clinical practice. Our study describes practical clinico-pathological specifications to optimize RAS ctDNA determination. Moreover, OncoBEAM™ is useful to monitor RAS in patients undergoing systemic therapy to detect resistance and evaluate the efficacy of particular treatments.
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            Protection from right- and left-sided colorectal neoplasms after colonoscopy: population-based study.

            Colonoscopy is used for early detection and prevention of colorectal cancer, but evidence on the magnitude of overall protection and protection according to anatomical site through colonoscopy performed in the community setting is sparse. We assessed whether receiving a colonoscopy in the preceding 10-year period, compared with no colonoscopy, was associated with prevalence of advanced colorectal neoplasms (defined as cancers or advanced adenomas) at various anatomical sites. A statewide cross-sectional study was conducted among 3287 participants in screening colonoscopy between May 1, 2005, and December 31, 2007, from the state of Saarland in Germany who were aged 55 years or older. Prevalence of advanced colorectal neoplasms was ascertained by screening colonoscopy and histopathologic examination of any polyps excised. Previous colonoscopy history was obtained by standardized questionnaire, and its association with prevalence of advanced colorectal neoplasms was estimated, after adjustment for potential confounding factors by log-binomial regression. Advanced colorectal neoplasms were detected in 308 (11.4%) of the 2701 participants with no previous colonoscopy compared with 36 (6.1%) of the 586 participants who had undergone colonoscopy within the preceding 10 years. After adjustment, overall and site-specific adjusted prevalence ratios for previous colonoscopy in the previous 10-year period were as follows: overall, 0.52 (95% confidence interval [CI] = 0.37 to 0.73); cecum and ascending colon, 0.99 (95% CI = 0.50 to 1.97); hepatic flexure and transverse colon, 1.21 (95% CI = 0.60 to 2.42); right-sided colon combined (cecum to transverse colon), 1.05 (95% CI = 0.63 to 1.76); splenic flexure and descending colon, 0.36 (95% CI = 0.16 to 0.82); sigmoid colon, 0.29 (95% CI = 0.16 to 0.53); rectum, 0.07 (95% CI = 0.02 to 0.40); left colon and rectum combined (splenic flexure to rectum, referred to as left-sided elsewhere), 0.33 (95% CI = 0.21 to 0.53). Prevalence of left-sided advanced colorectal neoplasms, but not right-sided advanced neoplasms, was strongly reduced within a 10-year period after colonoscopy, even in the community setting.
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              Point mutations of the N-ras gene in the blood plasma DNA of patients with myelodysplastic syndrome or acute myelogenous leukaemia.

              Oncogene mutations are frequently found in several tumour types and, among these, point mutations of the ras gene are particularly significant. A predominance of N-ras mutations has been found in the bone marrow DNA of patients with myelodysplastic syndrome (MDS) or acute myelogenous leukaemia (AML). On the other hand, increased levels of plasma DNA have previously been observed in patients suffering from various malignant diseases. In the present work we have investigated, by polymerase chain reaction (PCR), point mutations of the N-ras gene in the DNA of plasma, blood cells and bone marrow of 10 patients suffering from AML or MDS. The different ras mutations detected in five cases were always present in the plasma DNA while sometimes absent in the DNA of peripheral blood cells or bone marrow. This indicates that a bone marrow biopsy or aspiration does not necessarily contain all the malignant clones involved in the disease. Plasma could thus prove to be an easily accessible and useful material for detection and monitoring of myeloid disorders.
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                Author and article information

                Contributors
                Journal
                Biomed Res Int
                Biomed Res Int
                BMRI
                BioMed Research International
                Hindawi
                2314-6133
                2314-6141
                2020
                11 April 2020
                : 2020
                : 5419634
                Affiliations
                1Medical Oncology Department of Gastrointestinal Tumors, Liaoning Cancer Hospital & Institute, Cancer Hospital of China Medical University, No. 44, Xiaohe Road, Dadong District, Shenyang, 110042 Liaoning Province, China
                2Department of Medical Oncology, The First Hospital of China Medical University, Shenyang, China
                3Key Laboratory of Anticancer Drugs and Biotherapy of Liaoning Province, The First Hospital of China Medical University, Shenyang, China
                Author notes

                Guest Editor: Qiang Tong

                Author information
                https://orcid.org/0000-0001-9810-1368
                https://orcid.org/0000-0002-9645-7990
                https://orcid.org/0000-0002-0711-2764
                Article
                10.1155/2020/5419634
                7174943
                1d69446a-f1d1-45a6-a846-c163176012b2
                Copyright © 2020 Liuying Zhu et al.

                This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                : 22 July 2019
                : 21 March 2020
                Funding
                Funded by: Liaoning Cancer Hospital & Institute
                Award ID: Z1702
                Funded by: National Key R&D Program of China
                Award ID: #2018YFC1311600
                Funded by: Science and Technology Planning Project of Liaoning Province of China
                Award ID: 201800449
                Funded by: Science and Technology Planning Project of Shenyang
                Award ID: F15-139-9-27
                Award ID: 191124088
                Categories
                Research Article

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