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Topics {✒️}

quantitative real-time rt-pcr anti-transferrin receptor scfv-immunoliposomes open access article quantitative real-time pcr feline immunodeficiency virus virus-mimicking nanostructure system p53-mutant gastric cancer anti-transferrin receptor scfv tumor-suppressing signalling pathway tumor-derived cell lines pre-publication history β-galactosidase assay system mtt-based wst-1 assay cells' p53-mutant status article download pdf induces androgen-independent growth human gastric cancer p53 tumor-suppressing function jeffrey desano & liang xu control β-galactosidase plasmid lentiviral mir-34a infection mtt-based cytotoxicity assay mir-34a induced senescence gastric cancer cell full access human tumor regression clinical cancer research gastric cancer cells gastric cancer remains gastric cancer patients β-galactosidase activity detected mir-34 reporter assay renewal/differentiation decision-making bcl-2 3'utr reporter lentiviral packaging system natl cancer inst bcl-2 signaling pathway potential tumor suppressor cell stem cell ultra-low attachment plates tumor suppressor genes bcl-xl inhibition accompanied protects cancer cells tumor suppressor mir-34 tumor-suppressor mir-34 kato iii cells comprehensive cancer center reed jc regulating cell proliferation cell growth assay

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WebPage:
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         headline:Restoration of tumor suppressor miR-34 inhibits human p53-mutant gastric cancer tumorspheres
         description:MicroRNAs (miRNAs), some of which function as oncogenes or tumor suppressor genes, are involved in carcinogenesis via regulating cell proliferation and/or cell death. MicroRNA miR-34 was recently found to be a direct target of p53, functioning downstream of the p53 pathway as a tumor suppressor. miR-34 targets Notch, HMGA2, and Bcl-2, genes involved in the self-renewal and survival of cancer stem cells. The role of miR-34 in gastric cancer has not been reported previously. In this study, we examined the effects of miR-34 restoration on p53-mutant human gastric cancer cells and potential target gene expression. Human gastric cancer cells were transfected with miR-34 mimics or infected with the lentiviral miR-34-MIF expression system, and validated by miR-34 reporter assay using Bcl-2 3'UTR reporter. Potential target gene expression was assessed by Western blot for proteins, and by quantitative real-time RT-PCR for mRNAs. The effects of miR-34 restoration were assessed by cell growth assay, cell cycle analysis, caspase-3 activation, and cytotoxicity assay, as well as by tumorsphere formation and growth. Human gastric cancer Kato III cells with miR-34 restoration reduced the expression of target genes Bcl-2, Notch, and HMGA2. Bcl-2 3'UTR reporter assay showed that the transfected miR-34s were functional and confirmed that Bcl-2 is a direct target of miR-34. Restoration of miR-34 chemosensitized Kato III cells with a high level of Bcl-2, but not MKN-45 cells with a low level of Bcl-2. miR-34 impaired cell growth, accumulated the cells in G1 phase, increased caspase-3 activation, and, more significantly, inhibited tumorsphere formation and growth. Our results demonstrate that in p53-deficient human gastric cancer cells, restoration of functional miR-34 inhibits cell growth and induces chemosensitization and apoptosis, indicating that miR-34 may restore p53 function. Restoration of miR-34 inhibits tumorsphere formation and growth, which is reported to be correlated to the self-renewal of cancer stem cells. The mechanism of miR-34-mediated suppression of self-renewal appears to be related to the direct modulation of downstream targets Bcl-2, Notch, and HMGA2, indicating that miR-34 may be involved in gastric cancer stem cell self-renewal/differentiation decision-making. Our study suggests that restoration of the tumor suppressor miR-34 may provide a novel molecular therapy for p53-mutant gastric cancer.
         datePublished:2008-09-21T00:00:00Z
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            Gastric Cancer
            Cancer Stem Cell
            Gastric Cancer Cell
            Feline Immunodeficiency Virus
            Human Gastric Cancer Cell
            Cancer Research
            Oncology
            Surgical Oncology
            Health Promotion and Disease Prevention
            Biomedicine
            general
            Medicine/Public Health
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      headline:Restoration of tumor suppressor miR-34 inhibits human p53-mutant gastric cancer tumorspheres
      description:MicroRNAs (miRNAs), some of which function as oncogenes or tumor suppressor genes, are involved in carcinogenesis via regulating cell proliferation and/or cell death. MicroRNA miR-34 was recently found to be a direct target of p53, functioning downstream of the p53 pathway as a tumor suppressor. miR-34 targets Notch, HMGA2, and Bcl-2, genes involved in the self-renewal and survival of cancer stem cells. The role of miR-34 in gastric cancer has not been reported previously. In this study, we examined the effects of miR-34 restoration on p53-mutant human gastric cancer cells and potential target gene expression. Human gastric cancer cells were transfected with miR-34 mimics or infected with the lentiviral miR-34-MIF expression system, and validated by miR-34 reporter assay using Bcl-2 3'UTR reporter. Potential target gene expression was assessed by Western blot for proteins, and by quantitative real-time RT-PCR for mRNAs. The effects of miR-34 restoration were assessed by cell growth assay, cell cycle analysis, caspase-3 activation, and cytotoxicity assay, as well as by tumorsphere formation and growth. Human gastric cancer Kato III cells with miR-34 restoration reduced the expression of target genes Bcl-2, Notch, and HMGA2. Bcl-2 3'UTR reporter assay showed that the transfected miR-34s were functional and confirmed that Bcl-2 is a direct target of miR-34. Restoration of miR-34 chemosensitized Kato III cells with a high level of Bcl-2, but not MKN-45 cells with a low level of Bcl-2. miR-34 impaired cell growth, accumulated the cells in G1 phase, increased caspase-3 activation, and, more significantly, inhibited tumorsphere formation and growth. Our results demonstrate that in p53-deficient human gastric cancer cells, restoration of functional miR-34 inhibits cell growth and induces chemosensitization and apoptosis, indicating that miR-34 may restore p53 function. Restoration of miR-34 inhibits tumorsphere formation and growth, which is reported to be correlated to the self-renewal of cancer stem cells. The mechanism of miR-34-mediated suppression of self-renewal appears to be related to the direct modulation of downstream targets Bcl-2, Notch, and HMGA2, indicating that miR-34 may be involved in gastric cancer stem cell self-renewal/differentiation decision-making. Our study suggests that restoration of the tumor suppressor miR-34 may provide a novel molecular therapy for p53-mutant gastric cancer.
      datePublished:2008-09-21T00:00:00Z
      dateModified:2008-09-21T00:00:00Z
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      pageEnd:12
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      sameAs:https://doi.org/10.1186/1471-2407-8-266
      keywords:
         Gastric Cancer
         Cancer Stem Cell
         Gastric Cancer Cell
         Feline Immunodeficiency Virus
         Human Gastric Cancer Cell
         Cancer Research
         Oncology
         Surgical Oncology
         Health Promotion and Disease Prevention
         Biomedicine
         general
         Medicine/Public Health
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      name:Department of Radiation Oncology, Comprehensive Cancer Center, University of Michigan, Ann Arbor, USA
      name:Department of Radiation Oncology, Comprehensive Cancer Center, University of Michigan, Ann Arbor, USA
      name:Department of Radiation Oncology, Comprehensive Cancer Center, University of Michigan, Ann Arbor, USA
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      name:State Key Laboratory of Cancer Biology and Institute of Digestive Diseases, Xijing Hospital, Fourth Military Medical University, Xi'an, P.R. China
      name:Department of Radiation Oncology, Comprehensive Cancer Center, University of Michigan, Ann Arbor, USA

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