1. Academic Validation
  2. 1,25-dihydroxyvitamin D3 signaling-induced decreases in IRX4 inhibits NANOG-mediated cancer stem-like properties and gefitinib resistance in NSCLC cells

1,25-dihydroxyvitamin D3 signaling-induced decreases in IRX4 inhibits NANOG-mediated cancer stem-like properties and gefitinib resistance in NSCLC cells

  • Cell Death Dis. 2020 Aug 20;11(8):670. doi: 10.1038/s41419-020-02908-w.
Zhirong Jia  # 1 Yameng Zhang  # 1 Aiwen Yan  # 1 Meisa Wang 1 Qiushuang Han 1 Kaiwei Wang 1 Jie Wang 1 2 Chen Qiao 1 3 Zhenzhen Pan 1 Chuansheng Chen 1 Dong Hu 4 Xuansheng Ding 5 6
Affiliations

Affiliations

  • 1 School of Basic Medicine and Clinical Pharmacy, China Pharmaceutical University, 211198, Nanjing, China.
  • 2 Department of Pharmacy, the First Affiliated Hospital of Xinjiang Medical University, 830054, Urumqi, China.
  • 3 Precision Medicine Laboratory, School of Basic Medicine and Clinical Pharmacy, China Pharmaceutical University, 211198, Nanjing, China.
  • 4 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, 232001, Huainan, China. [email protected].
  • 5 School of Basic Medicine and Clinical Pharmacy, China Pharmaceutical University, 211198, Nanjing, China. [email protected].
  • 6 Precision Medicine Laboratory, School of Basic Medicine and Clinical Pharmacy, China Pharmaceutical University, 211198, Nanjing, China. [email protected].
  • # Contributed equally.
Abstract

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.

Figures
Products
  • Cat. No.
    Product Name
    Description
    Target
    Research Area
  • HY-17602
    98.07%, NANOG/CD133 Inhibitor