Cytotoxic Effect of p-Coumaric Acid on Neuroblastoma, N2a Cell via Generation of Reactive Oxygen Species Leading to Dysfunction of Mitochondria Inducing Apoptosis and Autophagy

• Published in: Molecular neurobiology Vol. 51; no. 1; pp. 119 - 130
• Main Authors: Shailasree, S., Venkataramana, M., Niranjana, S. R., Prakash, H. S.
• Format: Journal Article
• Language: English
• Published: Boston Springer US 01-02-2015; Springer Nature B.V
• Subjects: Animals; Apoptosis; Apoptosis - drug effects; Autophagy; Autophagy - drug effects; Biomarkers - metabolism; Biomedical and Life Sciences; Biomedicine; Caspase 8 - genetics; Caspase 8 - metabolism; Cell Biology; Cell Line, Tumor; Cell Shape - drug effects; Coumaric Acids - chemistry; Coumaric Acids - toxicity; Cytochromes c - metabolism; Cytotoxicity; Flow Cytometry; Gene Expression Regulation, Neoplastic - drug effects; Glutathione - metabolism; Membrane Potential, Mitochondrial - drug effects; Mice; Microtubule-Associated Proteins - metabolism; Mitochondria; Mitochondria - drug effects; Mitochondria - metabolism; Neurobiology; Neuroblastoma - enzymology; Neuroblastoma - genetics; Neuroblastoma - pathology; Neurology; Neurosciences; Propionates; Reactive Oxygen Species - metabolism; RNA, Messenger - genetics; RNA, Messenger - metabolism; Tumor Suppressor Protein p53 - genetics; Tumor Suppressor Protein p53 - metabolism; Up-Regulation - drug effects; Coumaric acid; Caspase-8; LC3-II; Neuroblastoma N2a cells; Autophagy; Apoptosis; p53
• ISSN: 0893-7648; 1559-1182
• DOI: 10.1007/s12035-014-8700-2

p
-Coumaric acid (
p
-CA), an ubiquitous plant phenolic acid, has been proven to render protection against pathological conditions. In the present study,
p
-CA was evaluated for its capacity to induce cytotoxic effect to neuroblastoma N2a cells and we report here the possible mechanism of its action.
p
-CA at a concentration of 150 μmol/L, upon exposure for 72 h, stimulated 81.23 % of cells to apoptosis, as evidenced by flow cytometer studies mediated through elevated levels of ROS (7.5-fold over control). Excess ROS production activated structural injury to mitochondrial membrane, observed as dissipation of its membrane potential and followed by the release of cytochrome
c
(8.73-fold). Enhanced generation of intracellular ROS correlated well with the decreased levels (~60 %) of intracellular GSH. Sensitizing neuroblastoma cells for induction of apoptosis by
p
-CA identified p53-mediated upregulated accumulation of caspase-8 messenger RNA (2.8-fold). Our data report on autophagy, representing an additional mechanism of
p
-CA to induce growth arrest, detected by immunoblotting and fluorescence, correlated with accumulation of elevated levels (1.2-fold) of the LC3-II protein and acridine orange-stained autophagosomes, both autophagy markers. The present study indicates
p
-CA was effective in production of ROS-dependent mitochondrial damage-induced cytotoxicity in N2a cells.