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nature portfolio beta-catenin-mediated epithelial-mesenchymal transition vdr/plc-gamma1/tgf-beta1/pathway serum‐free opti‐mem media research groups privacy policy tgf-β/smad signaling-induced irx4 tgf-β1/smad3 signaling pathway tgf-β/smad signaling pathway28 nature small-molecule multi-kinase inhibitor20 advertising erk1/2-mzf1 signaling pathway reprints tgf-β1/smad3 signaling japan activating stat3/cxcl1 axis 2d3 signaling-induced decreases egfr-tyrosine kinase inhibitors small-cell lung cancer egfr-tki resistant cells irx4-mediated cancer stem potentiates cell death social media poor egfr-tki sensitivity inhibit egfr-tki resistance irx4-expressing cell lines nanog-mediated stem cell state key laboratory regulating egfr-tki resistance cd44+cd133+ pc-9/gr cells increase egfr-tki resistance high tgf-β1 levels irx-family genes participate egfr-mutant nsclc cells9 contract number hh20190622lhy-lv03 ventricular-specific progenitor cell multi-directional differentiation potential pnanog-luc reporter activity irx4+mouse embryonic cells original author tgf-β1 induced permissions sirna/plasmid‐lipid complexes lung cancer progression tyrosine kinase inhibitors 20 ng/ml tgf-β1 delayed development cell death dis pre-clinical models support

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• How IRX4 controls gefitinib cytotoxicity?
• How to regulate IRX4-mediated cancer stem-like properties and gefitinib resistance?

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         headline:1,25-dihydroxyvitamin D3 signaling-induced decreases in IRX4 inhibits NANOG-mediated cancer stem-like properties and gefitinib resistance in NSCLC cells
         description:Recent studies have demonstrated that acquisition of cancer stem-like properties plays an essential role in promoting epidermal growth factor receptor-tyrosine kinase inhibitors (EGFR-TKIs) resistance in non-small cell lung cancer (NSCLC); however, how to regulate cancer stem-like properties and EGFR-TKI resistance is largely unclear. In this study, we discovered that increased iroquois-class homeodomain protein 4 (IRX4) was related to gefitinib resistance in NSCLC cells. Knockdown of IRX4 inhibited cell proliferation, sphere formation, and the expression of CD133, ALDH1A1, NANOG, Sox2 and Notch1, and the transcriptional activity of NANOG promoter. IRX4 overexpression increased the protein level of NANOG and CD133 in PC-9 cells. Combination of knocking-down IRX4 with gefitinib increased cell apoptosis and decreased cell viability and the expression of p-EGFR and NANOG in PC-9/GR cells. IRX4 knockdown in a PC-9/GR xenograft tumor model inhibited tumor progression and the expression of NANOG and CD133 more effectively than single treatment alone. Knockdown of NANOG inhibited the expression of CD133 and restored gefitinib cytotoxicity, and NANOG overexpression-induced cancer stem-like properties and gefitinib resistance could be obviously reversed by knocking-down IRX4. Further, we found that 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) reduced obviously the expression of IRX4 and NANOG by inhibiting the activation of TGF-β1/Smad3 signaling pathway; moreover, combination of 1,25(OH)2D3 and gefitinib decreased cell viability and proliferation or tumor progression and the expression of IRX4 and NANOG compared with single treatment alone both in PC-9/GR cells and in a PC-9/GR xenograft tumor model. These results reveal that inhibition of IRX4-mediated cancer stem-like properties by regulating 1,25(OH)2D3 signaling may increase gefitinib cytotoxicity. Combination therapy of gefitinib and 1,25(OH)2D3 by targeting IRX4 and NANOG, could provide a promising strategy to improve gefitinib cytotoxicity.
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               type:PostalAddress
            type:Organization
      name:Zhenzhen Pan
      affiliation:
            name:China Pharmaceutical University
            address:
               name:School of Basic Medicine and Clinical Pharmacy, China Pharmaceutical University, Nanjing, China
               type:PostalAddress
            type:Organization
      name:Chuansheng Chen
      affiliation:
            name:China Pharmaceutical University
            address:
               name:School of Basic Medicine and Clinical Pharmacy, China Pharmaceutical University, Nanjing, China
               type:PostalAddress
            type:Organization
      name:Dong Hu
      affiliation:
            name:Anhui University of Science and Technology
            address:
               name:Key Laboratory of Industrial Dust Prevention and Control & Occupational Safety and Health of the Ministry of Education, Medical School, Anhui University of Science and Technology, Huainan, China
               type:PostalAddress
            type:Organization
      email:[email protected]
      name:Xuansheng Ding
      affiliation:
            name:China Pharmaceutical University
            address:
               name:School of Basic Medicine and Clinical Pharmacy, China Pharmaceutical University, Nanjing, China
               type:PostalAddress
            type:Organization
            name:China Pharmaceutical University
            address:
               name:Precision Medicine Laboratory, School of Basic Medicine and Clinical Pharmacy, China Pharmaceutical University, Nanjing, China
               type:PostalAddress
            type:Organization
      email:[email protected]
PostalAddress:
      name:School of Basic Medicine and Clinical Pharmacy, China Pharmaceutical University, Nanjing, China
      name:School of Basic Medicine and Clinical Pharmacy, China Pharmaceutical University, Nanjing, China
      name:School of Basic Medicine and Clinical Pharmacy, China Pharmaceutical University, Nanjing, China
      name:School of Basic Medicine and Clinical Pharmacy, China Pharmaceutical University, Nanjing, China
      name:School of Basic Medicine and Clinical Pharmacy, China Pharmaceutical University, Nanjing, China
      name:School of Basic Medicine and Clinical Pharmacy, China Pharmaceutical University, Nanjing, China
      name:School of Basic Medicine and Clinical Pharmacy, China Pharmaceutical University, Nanjing, China
      name:Department of Pharmacy, the First Affiliated Hospital of Xinjiang Medical University, Urumqi, China
      name:School of Basic Medicine and Clinical Pharmacy, China Pharmaceutical University, Nanjing, China
      name:Precision Medicine Laboratory, School of Basic Medicine and Clinical Pharmacy, China Pharmaceutical University, Nanjing, China
      name:School of Basic Medicine and Clinical Pharmacy, China Pharmaceutical University, Nanjing, China
      name:School of Basic Medicine and Clinical Pharmacy, China Pharmaceutical University, Nanjing, China
      name:Key Laboratory of Industrial Dust Prevention and Control & Occupational Safety and Health of the Ministry of Education, Medical School, Anhui University of Science and Technology, Huainan, China
      name:School of Basic Medicine and Clinical Pharmacy, China Pharmaceutical University, Nanjing, China
      name:Precision Medicine Laboratory, School of Basic Medicine and Clinical Pharmacy, China Pharmaceutical University, Nanjing, China

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