Digitoxin Affects Metabolism, ROS Production and Proliferation in Pancreatic Cancer Cells Differently Depending on the Cell Phenotype

Abstract

Digitoxin has repeatedly shown to have negative effects on cancer cell viability; however, the actual mechanism is still unknown. In this study, we investigated the effects of digitoxin (1–100 nM) in four pancreatic cancer cell lines, BxPC-3, CFPAC-1, Panc-1, and AsPC-1. The cell lines differ in their KRAS/BRAF mutational status and primary tumor or metastasis origin. We could detect differences in the basal rates of cell proliferation, glycolysis, and ROS production, giving the cell lines different phenotypes. Digitoxin treatment induced apoptosis in all four cell lines, but to different degrees. Cells derived from primary tumors (Panc-1 and BxPC-3) were highly proliferating with a high proportion of cells in the S/G2 phase, and were more sensitive to digitoxin treatment than the cell lines derived from metastases (CFPAC-1 and AsPC-1), with a high proportion of cells in G0/G1. In addition, the effects of digitoxin on the rate of glycolysis, ROS production, and proliferation were dependent on the basal metabolism and origin of the cells. The KRAS downstream signaling pathways were not altered by digitoxin treatment, thus the effects exerted by digitoxin were probably disconnected from these signaling pathways. We conclude that digitoxin is a promising treatment in highly proliferating pancreatic tumors.

Cytotoxic and Pro-Apoptotic Effects of Leaves Extract of Antiaris africana Engler (Moraceae)

“…Based on these results, the presence of toxicarioside K and antiaroside ZC, or other unidentified CGs, in the leaf extract of Antiaris africana Engler may also explain the cytotoxicity induced for THP-1, ASPC-1, SW602 and MCF-7 cells lines observed here. It has been proven that CGs exhibit anti-proliferative activity by inhibiting important signaling pathways, such as KRAS, that regulate cell growth, and are known to induce the mitochondrial apoptotic pathway via ROS-mediated oxidative stress [35][36][37]. Therefore, we expected that similar molecular mechanisms could implicate in EHA-Aq-triggered apoptotic cell death.…”

Incorporating Network Pharmacology and Experimental Validation to Identify Bioactive Compounds and Potential Mechanisms of Digitalis in Treating Anaplastic Thyroid Cancer

“… 54 Astragaloside A can inhibit the progression of colorectal cancer cells 55 and inhibit the migration and EMT progression of cervical cancer cells. 56 Digitoxin has an inhibitory effect on pancreatic cancer cells 57 and can reduce HIF-1α and STAT3 to promote apoptosis and inhibit the proliferation and migration of KRAS mutant human colon cancer cells. 58 However, the effects of these active ingredients on thyroid cancer, especially undifferentiated thyroid cancer, have not yet been reported.…”

Exosomal SOX21-AS1 Regulates EREG by Sponging miR-451a and Promotes the Malignancy of Pancreatic Ductal Adenocarcinoma

“…The normal human pancreatic ductal cell line HPDE and pancreatic adenocarcinoma cell lines CFPAC-1, AsPC-1, BxPc-3, PANC-1 and MIA PaCa-2 were purchased from the American Type Culture Collection (ATCC, Manassas, VA) and incubated in high glucose Dulbecco's Modified Eagle Medium (DMEM) (Invitrogen, Thermo Fisher Scientific, Inc., Carlsbad, CA, USA) supplemented with 1% penicillin/streptomycin (Invitrogen), 10% foetal bovine serum (FBS; Gibco, Life Technologies, Carlsbad, CA, USA) and 25 mmol/L HEPES (Thermo Fisher, Dreieich, Germany) at 37 °C in a humidified atmosphere with 5% CO2. HUVECs (Institute of Cell Biology of the Chinese Academy of Sciences, Shanghai, China) were cultured in MCDB 131 medium (GIBCO) supplemented with 5% microvascular growth supplement (GIBCO) and 1% GlutaMAX (GIBCO) under the same conditions as those used for PDAC 24 , 25 . Lentiviruses overexpressing SOX21-AS1 (SOX21-AS1), a small hairpin RNA (shRNA) against SOX21-AS1 (shSOX21-AS1), miR mimic, miR inhibitor, EREG overexpression vector (EREG), shRNA targeting EREG (shEREG) and the corresponding NCs (NC, shNC, mimic NC, inhibitor NC, EREG NC and shEREG NC) were procured from GenePharma (Shanghai, China).…”