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

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) 2 D 3) reduced obviously the expression of IRX4 and NANOG by inhibiting the activation of TGF-β1/Smad3 signaling pathway; moreover, combination of 1,25(OH) 2 D 3 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) 2 D 3 signaling may increase gefitinib cytotoxicity. Combination therapy of gefitinib and 1,25 (OH) 2 D 3 by targeting IRX4 and NANOG, could provide a promising strategy to improve gefitinib cytotoxicity.

Antitumoral Effects of Dovitinib in Triple-Negative Breast Cancer are Synergized by Calcitriol in Vivo and In Vitro

“…Similarly, previous studies have shown that vitamin D derivatives exert synergistic effects when used in combination with other oncological drugs [18][19][20][21]. In particular, it has been demonstrated that the combination of calcitriol or its analogs with gefitinib, a synthetic RTKI, enhances global anticancer activity by inhibiting tumor growth and inducing apoptosis, and in cancer patients it does not result in serious undesirable side effects [17,20,43,44]. To the best of our knowledge, this is the first study addressing the antineoplastic effects of dovitinib in combination with calcitriol.…”

“…Calcitriol, the vitamin D most active metabolite, exert potent antineoplastic activity by modulating diverse signaling networks involved in inhibition of cell proliferation, anti-inflammatory effects, acquisition of a more differentiated phenotype and induction of apoptosis [16]. Moreover, calcitriol has been shown to increase the sensitivity of tumor cells to various chemotherapeutic agents [17][18][19][20][21], with the added benefit of being a natural compound derived from dietary sources or by sun exposure. Notably, low vitamin D serum levels have been shown to correlate with increased risk of certain neoplasms, including breast cancer [22].…”

Wnt Activity and Cell Proliferation Are Coupled to Extracellular Vesicle Release in Multiple Organoid Models

“…Intra-tumoral cellular heterogeneity is a major hallmark of lung cancers. Several molecules, such as CD133 ( Chen et al, 2008 ; Jia et al, 2020 ), have been suggested as markers of the aggressive cancer cell population with stem cell features. Importantly, although PORCN+ cells have a critical role in LUAD progression in a mouse model ( Tammela et al, 2017 ), no attempt has been made to isolate these Wnt-producing niche cells.…”

Identification of Targetable Liabilities in the Dynamic Metabolic Profile of EGFR-Mutant Lung Adenocarcinoma: Thinking beyond Genomics for Overcoming EGFR TKI Resistance

“…Dichloroacetate PDHK inhibitor Induction of Acetyl-CoA formation [26] Atorvastatin CAV-1/GLUT-3 inhibitor Inhibition of glucose uptake and cholesterol synthesis [30] PF-429242 SREBP1 inhibitor Inhibition of lipid synthesis [31] Orlistat FASN inhibitor Inhibition of EGFR palmitoylation [32] Epalrestat AKR1B1 inhibitor Inhibition of GSH synthesis [33] PiperlongumineAuranofin ROS inducing agent Induction of oxidative stress [34] Buthionine sulfoximine GSH synthesis inhibitor Inhibition of GSH synthesis [34] 1,25D VDR agonist Inhibition of stemness phenotype [35] AZ12756122 FASN inhibitor Inhibition of stemness phenotype [36] Abbreviations: PDHK, pyruvate dehydrogenase kinase; Acetyl-CoA, acetyl coenzyme A; CAV-1, caveolin-1; GLUT-3, glucose transporter-3; FASN, fatty acid synthase; EGFR, epidermal growth factor receptor; SREBP1, sterol regulatory-element-binding protein 1; AKR1B1, aldo-keto reductase family 1 member B1; GSH, glutathione; ROS, reactive oxygen species; 1,25D, 1,25-dihydroxyvitamin D3; VDR, vitamin D receptor.…”