Pantoprazole Induces Mitochondrial Apoptosis and Attenuates NF-κB Signaling in Glioma Cells
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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.
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