Pantoprazole Induces Mitochondrial Apoptosis and Attenuates NF-κB Signaling in Glioma Cells
- 216 Downloads
Gastric H+/K+-ATPase or vacuolar-ATPases (V-ATPases) are critical for the cancer cells survival and growth in the ischemic microenvironment by extruding protons from the cell. The drugs which inhibit V-ATPases are known as proton pump inhibitors (PPIs). In the present study, we aimed to evaluate the anticancer efficacy of pantoprazole (PPZ) and its consequences on NF-κB signaling in glioma cells. We have used MTT and clonogenic assay to show PPZ effect on glioma cell growth. Propidium iodide and rhodamine 123 staining were performed to demonstrate cell cycle arrest and mitochondrial depolarization. TUNEL staining was used to evidence apoptosis after PPZ treatment. Immunoblotting and immunofluorescence microscopy were performed to depict protein levels and localization, respectively. Luciferase assay was performed to confirm NF-κB suppression by PPZ. Our results revealed PPZ treatment inhibits cell viability or growth and induced cell death in a dose- and time-dependent manner. PPZ exposure arrested G0/G1 cyclic phase and increased TUNEL positivity, caspase-3 and PARP cleavage with altered pro and anti-apoptotic proteins. PPZ also induced ROS levels and depolarized mitochondria (Δψm) with increased cytosolic cytochrome c level. Further, PPZ suppressed TNF-α stimulated NF-κB signaling by repressing p65 nuclear translocation. NF-κB luciferase reporter assays revealed significant inhibition of NF-κB gene upon PPZ treatment. PPZ exposure also reduced the expression of NF-κB-associated genes, such as cyclin-D1, iNOS, and COX-2, which indicate NF-κB inhibition. Altogether, the present study disclosed that PPZ exerts mitochondrial apoptosis and attenuates NF-κB signaling suggesting PPZ can be an effective and safe anticancer drug for glioma.
KeywordsGlioma V-ATPases Pantoprazole Anticancer Apoptosis NF-κB signaling
We acknowledge Ms. Nalini M. (Microscopic facility, University of Hyderabad) for the technical assistance of confocal microscopy.
KV, DB, and PPB are responsible for the conceptualization and design of the studies. KV and DB performed all experiments and analyzed the data. KV and DB wrote the manuscript. KV, DB, and PPB reviewed and finalized the final draft.
We acknowledge University with potential for excellence (UPE) India (UH/UGC/UPE-2 Interface studies/Research Projects/B1.4) and Department of Science and technology (DST-India) (SB/EMEQ-257/2013) for lab funding. KG acknowledges financial support from Council of Scientific and Industrial Research (CSIR), India (Award No. F.No:09/414(0863)/2009-EMR-I). DB is thankful to the Department of Biotechnology (DBT), India (Award No. DBT/JRF/13/AL/132/2043) for student fellowship.
Compliance with Ethical Standards
Conflict of interest
The authors declare no conflicts of interest.
- Breedveld P, Pluim D, Cipriani G, Wielinga P, van Tellingen O, Schinkel AH, Schellens JH (2005) The effect of Bcrp1 (Abcg2) on the in vivo pharmacokinetics and brain penetration of imatinib mesylate (Gleevec): implications for the use of breast cancer resistance protein and P-glycoprotein inhibitors to enable the brain penetration of imatinib in patients. Cancer Res 65(7):2577–2582. https://doi.org/10.1158/0008-5472.CAN-04-2416 CrossRefPubMedGoogle Scholar
- Chen M, Huang SL, Zhang XQ, Zhang B, Zhu H, Yang VW, Zou XP (2012) Reversal effects of pantoprazole on multidrug resistance in human gastric adenocarcinoma cells by down-regulating the V-ATPases/mTOR/HIF-1alpha/P-gp and MRP1 signaling pathway in vitro and in vivo. J Cell Biochem 113(7):2474–2487. https://doi.org/10.1002/jcb.24122 CrossRefPubMedPubMedCentralGoogle Scholar
- De Milito A, Iessi E, Logozzi M, Lozupone F, Spada M, Marino ML, Federici C, Perdicchio M, Matarrese P, Lugini L, Nilsson A, Fais S (2007) Proton pump inhibitors induce apoptosis of human B-cell tumors through a caspase-independent mechanism involving reactive oxygen species. Cancer Res 67(11):5408–5417. https://doi.org/10.1158/0008-5472.CAN-06-4095 CrossRefPubMedGoogle Scholar
- Du L, Lyle CS, Obey TB, Gaarde WA, Muir JA, Bennett BL, Chambers TC (2004) Inhibition of cell proliferation and cell cycle progression by specific inhibition of basal JNK activity: evidence that mitotic Bcl-2 phosphorylation is JNK-independent. J Biol Chem 279(12):11957–11966. https://doi.org/10.1074/jbc.M304935200 CrossRefPubMedGoogle Scholar
- Ji BC, Yu CC, Yang ST, Hsia TC, Yang JS, Lai KC, Ko YC, Lin JJ, Lai TY, Chung JG (2012) Induction of DNA damage by deguelin is mediated through reducing DNA repair genes in human non-small cell lung cancer NCI-H460 cells. Oncol Rep 27(4):959–964. https://doi.org/10.3892/or.2012.1622 CrossRefPubMedPubMedCentralGoogle Scholar
- Louis DN (2006) Molecular pathology of malignant gliomas. Annu Rev Pathol 1:97–117. https://doi.org/10.1146/annurev.pathol.1.110304.100043 CrossRefPubMedGoogle Scholar
- Luciani F, Spada M, De Milito A, Molinari A, Rivoltini L, Montinaro A, Marra M, Lugini L, Logozzi M, Lozupone F, Federici C, Iessi E, Parmiani G, Arancia G, Belardelli F, Fais S (2004) Effect of proton pump inhibitor pretreatment on resistance of solid tumors to cytotoxic drugs. J Natl Cancer Inst 96(22):1702–1713. https://doi.org/10.1093/jnci/djh305 CrossRefPubMedGoogle Scholar
- Martin de Argila C (2005) Safety of potent gastric acid inhibition. Drugs 65 Suppl 1:97–104Google Scholar
- Neuwelt EA, Bauer B, Fahlke C, Fricker G, Iadecola C, Janigro D, Leybaert L, Molnar Z, O’Donnell ME, Povlishock JT, Saunders NR, Sharp F, Stanimirovic D, Watts RJ, Drewes LR (2011) Engaging neuroscience to advance translational research in brain barrier biology. Nat Rev Neurosci 12(3):169–182. https://doi.org/10.1038/nrn2995 CrossRefPubMedPubMedCentralGoogle Scholar
- Ostrom QT, Gittleman H, Fulop J, Liu M, Blanda R, Kromer C, Wolinsky Y, Kruchko C, Barnholtz-Sloan JS (2015) CBTRUS statistical report: primary brain and central nervous system tumors diagnosed in the United States in 2008–2012. Neuro Oncol 17(Suppl 4):iv1–iv62. https://doi.org/10.1093/neuonc/nov189 CrossRefPubMedPubMedCentralGoogle Scholar
- Portnow J, Badie B, Chen M, Liu A, Blanchard S, Synold TW (2009) The neuropharmacokinetics of temozolomide in patients with resectable brain tumors: potential implications for the current approach to chemoradiation. Clin Cancer Res 15(22):7092–7098. https://doi.org/10.1158/1078-0432.CCR-09-1349 CrossRefPubMedPubMedCentralGoogle Scholar
- Sigaroudi A, Stelzer C, Braun T, Frechen S, Huttner S, Schroter M, Kinzig M, Fuhr U, Holzgrabe U, Sorgel F (2016) Comparison of pantoprazole concentrations in simultaneous cerebrospinal fluid and serum samples. Pharmacology 98(1–2):70–72. https://doi.org/10.1159/000445720 CrossRefPubMedGoogle Scholar
- Tanigawa T, Watanabe T, Higuchi K, Machida H, Okazaki H, Yamagami H, Watanabe K, Tominaga K, Fujiwara Y, Oshitani N, Arakawa T (2009) Lansoprazole, a proton pump inhibitor, suppresses production of tumor necrosis factor-alpha and interleukin-1beta induced by lipopolysaccharide and helicobacter pylori bacterial components in human monocytic cells via inhibition of activation of nuclear factor-kappaB and extracellular signal-regulated kinase. J Clin Biochem Nutr 45(1):86–92. https://doi.org/10.3164/jcbn.08-267 CrossRefPubMedPubMedCentralGoogle Scholar
- Udelnow A, Kreyes A, Ellinger S, Landfester K, Walther P, Klapperstueck T, Wohlrab J, Henne-Bruns D, Knippschild U, Wurl P (2011) Omeprazole inhibits proliferation and modulates autophagy in pancreatic cancer cells. PLoS ONE 6(5):e20143. https://doi.org/10.1371/journal.pone.0020143 CrossRefPubMedPubMedCentralGoogle Scholar