MicroRNA-224 is associated with colorectal cancer progression and response to 5-fluorouracil-based chemotherapy by KRAS-dependent and-independent mechanisms

E. B. Amankwatia, P. Chakravarty, F. A. Carey, S. Weidlich, R. J. C. Steele, A. J. Munro, C. R. Wolf, G. Smith (Lead / Corresponding author)

    Research output: Contribution to journalArticle

    42 Citations (Scopus)

    Abstract

    Background:Colorectal cancers arise from benign adenomas, although not all adenomas progress to cancer and there are marked interpatient differences in disease progression. We have previously associated KRAS mutations with disease progression and reduced survival in colorectal cancer patients.Methods:We used TaqMan low-density array (TLDA) qRT-PCR analysis to identify miRNAs differentially expressed in normal colorectal mucosa, adenomas and cancers and in isogeneic KRAS WT and mutant HCT116 cells, and used a variety of phenotypic assays to assess the influence of miRNA expression on KRAS activity, chemosensitivity, proliferation and invasion.Results:MicroRNA-224 was differentially expressed in dysplastic colorectal disease and in isogeneic KRAS WT and mutant HCT116 cells. Antagomir-mediated miR-224 silencing in HCT116 KRAS WT cells phenocopied KRAS mutation, increased KRAS activity and ERK and AKT phosphorylation. 5-FU chemosensitivity was significantly increased in miR-224 knockdown cells, and in NIH3T3 cells expressing KRAS and BRAF mutant proteins. Bioinformatics analysis of predicted miR-224 target genes predicted altered cell proliferation, invasion and epithelial-mesenchymal transition (EMT) phenotypes that were experimentally confirmed in miR-224 knockdown cells.Conclusions:We describe a novel mechanism of KRAS regulation, and highlight the clinical utility of colorectal cancer-specific miRNAs as disease progression or clinical response biomarkers.

    Original languageEnglish
    Pages (from-to)1480-1490
    Number of pages11
    JournalBritish Journal of Cancer
    Volume112
    Issue number9
    DOIs
    Publication statusPublished - 28 Apr 2015

    Fingerprint

    MicroRNAs
    Fluorouracil
    Colorectal Neoplasms
    Adenoma
    HCT116 Cells
    Disease Progression
    Drug Therapy
    Mutation
    Epithelial-Mesenchymal Transition
    Mutant Proteins
    Computational Biology
    Neoplasms
    Mucous Membrane
    Biomarkers
    Phosphorylation
    Cell Proliferation
    Phenotype
    Polymerase Chain Reaction
    Survival
    Genes

    Keywords

    • Chemotherapy
    • Colorectal adenoma
    • Colorectal cancer
    • KRAS
    • microRNA
    • miR-224

    Cite this

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    title = "MicroRNA-224 is associated with colorectal cancer progression and response to 5-fluorouracil-based chemotherapy by KRAS-dependent and-independent mechanisms",
    abstract = "Background:Colorectal cancers arise from benign adenomas, although not all adenomas progress to cancer and there are marked interpatient differences in disease progression. We have previously associated KRAS mutations with disease progression and reduced survival in colorectal cancer patients.Methods:We used TaqMan low-density array (TLDA) qRT-PCR analysis to identify miRNAs differentially expressed in normal colorectal mucosa, adenomas and cancers and in isogeneic KRAS WT and mutant HCT116 cells, and used a variety of phenotypic assays to assess the influence of miRNA expression on KRAS activity, chemosensitivity, proliferation and invasion.Results:MicroRNA-224 was differentially expressed in dysplastic colorectal disease and in isogeneic KRAS WT and mutant HCT116 cells. Antagomir-mediated miR-224 silencing in HCT116 KRAS WT cells phenocopied KRAS mutation, increased KRAS activity and ERK and AKT phosphorylation. 5-FU chemosensitivity was significantly increased in miR-224 knockdown cells, and in NIH3T3 cells expressing KRAS and BRAF mutant proteins. Bioinformatics analysis of predicted miR-224 target genes predicted altered cell proliferation, invasion and epithelial-mesenchymal transition (EMT) phenotypes that were experimentally confirmed in miR-224 knockdown cells.Conclusions:We describe a novel mechanism of KRAS regulation, and highlight the clinical utility of colorectal cancer-specific miRNAs as disease progression or clinical response biomarkers.",
    keywords = "Chemotherapy, Colorectal adenoma, Colorectal cancer, KRAS, microRNA, miR-224",
    author = "Amankwatia, {E. B.} and P. Chakravarty and Carey, {F. A.} and S. Weidlich and Steele, {R. J. C.} and Munro, {A. J.} and Wolf, {C. R.} and G. Smith",
    year = "2015",
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    day = "28",
    doi = "10.1038/bjc.2015.125",
    language = "English",
    volume = "112",
    pages = "1480--1490",
    journal = "British Journal of Cancer",
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    publisher = "Cancer Research UK",
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    }

    MicroRNA-224 is associated with colorectal cancer progression and response to 5-fluorouracil-based chemotherapy by KRAS-dependent and-independent mechanisms. / Amankwatia, E. B.; Chakravarty, P.; Carey, F. A.; Weidlich, S.; Steele, R. J. C.; Munro, A. J.; Wolf, C. R.; Smith, G. (Lead / Corresponding author).

    In: British Journal of Cancer, Vol. 112, No. 9, 28.04.2015, p. 1480-1490.

    Research output: Contribution to journalArticle

    TY - JOUR

    T1 - MicroRNA-224 is associated with colorectal cancer progression and response to 5-fluorouracil-based chemotherapy by KRAS-dependent and-independent mechanisms

    AU - Amankwatia, E. B.

    AU - Chakravarty, P.

    AU - Carey, F. A.

    AU - Weidlich, S.

    AU - Steele, R. J. C.

    AU - Munro, A. J.

    AU - Wolf, C. R.

    AU - Smith, G.

    PY - 2015/4/28

    Y1 - 2015/4/28

    N2 - Background:Colorectal cancers arise from benign adenomas, although not all adenomas progress to cancer and there are marked interpatient differences in disease progression. We have previously associated KRAS mutations with disease progression and reduced survival in colorectal cancer patients.Methods:We used TaqMan low-density array (TLDA) qRT-PCR analysis to identify miRNAs differentially expressed in normal colorectal mucosa, adenomas and cancers and in isogeneic KRAS WT and mutant HCT116 cells, and used a variety of phenotypic assays to assess the influence of miRNA expression on KRAS activity, chemosensitivity, proliferation and invasion.Results:MicroRNA-224 was differentially expressed in dysplastic colorectal disease and in isogeneic KRAS WT and mutant HCT116 cells. Antagomir-mediated miR-224 silencing in HCT116 KRAS WT cells phenocopied KRAS mutation, increased KRAS activity and ERK and AKT phosphorylation. 5-FU chemosensitivity was significantly increased in miR-224 knockdown cells, and in NIH3T3 cells expressing KRAS and BRAF mutant proteins. Bioinformatics analysis of predicted miR-224 target genes predicted altered cell proliferation, invasion and epithelial-mesenchymal transition (EMT) phenotypes that were experimentally confirmed in miR-224 knockdown cells.Conclusions:We describe a novel mechanism of KRAS regulation, and highlight the clinical utility of colorectal cancer-specific miRNAs as disease progression or clinical response biomarkers.

    AB - Background:Colorectal cancers arise from benign adenomas, although not all adenomas progress to cancer and there are marked interpatient differences in disease progression. We have previously associated KRAS mutations with disease progression and reduced survival in colorectal cancer patients.Methods:We used TaqMan low-density array (TLDA) qRT-PCR analysis to identify miRNAs differentially expressed in normal colorectal mucosa, adenomas and cancers and in isogeneic KRAS WT and mutant HCT116 cells, and used a variety of phenotypic assays to assess the influence of miRNA expression on KRAS activity, chemosensitivity, proliferation and invasion.Results:MicroRNA-224 was differentially expressed in dysplastic colorectal disease and in isogeneic KRAS WT and mutant HCT116 cells. Antagomir-mediated miR-224 silencing in HCT116 KRAS WT cells phenocopied KRAS mutation, increased KRAS activity and ERK and AKT phosphorylation. 5-FU chemosensitivity was significantly increased in miR-224 knockdown cells, and in NIH3T3 cells expressing KRAS and BRAF mutant proteins. Bioinformatics analysis of predicted miR-224 target genes predicted altered cell proliferation, invasion and epithelial-mesenchymal transition (EMT) phenotypes that were experimentally confirmed in miR-224 knockdown cells.Conclusions:We describe a novel mechanism of KRAS regulation, and highlight the clinical utility of colorectal cancer-specific miRNAs as disease progression or clinical response biomarkers.

    KW - Chemotherapy

    KW - Colorectal adenoma

    KW - Colorectal cancer

    KW - KRAS

    KW - microRNA

    KW - miR-224

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    DO - 10.1038/bjc.2015.125

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    EP - 1490

    JO - British Journal of Cancer

    JF - British Journal of Cancer

    SN - 0007-0920

    IS - 9

    ER -