Limits...
Induction of apoptosis and antiproliferative activity of naringenin in human epidermoid carcinoma cell through ROS generation and cell cycle arrest.

Ahamad MS, Siddiqui S, Jafri A, Ahmad S, Afzal M, Arshad M - PLoS ONE (2014)

Bottom Line: The search for antiproliferative agents that reduce skin carcinoma is a task of great importance.It also caused naringenin-mediated epidermoid carcinoma apoptosis by inducing mitochondrial depolarization.Cell cycle study showed that naringenin induced cell cycle arrest in G0/G1 phase of cell cycle and caspase-3 analysis revealed a dose dependent increment in caspase-3 activity which led to cell apoptosis.

View Article: PubMed Central - PubMed

Affiliation: Department of Zoology, Shibli National (PG) College, Azamgarh, Uttar Pradesh, India.

ABSTRACT
A natural predominant flavanone naringenin, especially abundant in citrus fruits, has a wide range of pharmacological activities. The search for antiproliferative agents that reduce skin carcinoma is a task of great importance. The objective of this study was to analyze the anti-proliferative and apoptotic mechanism of naringenin using MTT assay, DNA fragmentation, nuclear condensation, change in mitochondrial membrane potential, cell cycle kinetics and caspase-3 as biomarkers and to investigate the ability to induce reactive oxygen species (ROS) initiating apoptotic cascade in human epidermoid carcinoma A431 cells. Results showed that naringenin exposure significantly reduced the cell viability of A431 cells (p<0.01) with a concomitant increase in nuclear condensation and DNA fragmentation in a dose dependent manner. The intracellular ROS generation assay showed statistically significant (p<0.001) dose-related increment in ROS production for naringenin. It also caused naringenin-mediated epidermoid carcinoma apoptosis by inducing mitochondrial depolarization. Cell cycle study showed that naringenin induced cell cycle arrest in G0/G1 phase of cell cycle and caspase-3 analysis revealed a dose dependent increment in caspase-3 activity which led to cell apoptosis. This study confirms the efficacy of naringenin that lead to cell death in epidermoid carcinoma cells via inducing ROS generation, mitochondrial depolarization, nuclear condensation, DNA fragmentation, cell cycle arrest in G0/G1 phase and caspase-3 activation.

Show MeSH

Related in: MedlinePlus

Effect of naringenin on different phases of cell cycle.A431 cells were treated with different concentrations of naringenin (100 µM−300 µM) for 24 h, stained with propidium iodide and measured by flow cytometry (A) Representative photomicrograph showing the apoptosis and phase distribution of cell population (B) Quantitative distribution of percentage of A431 cells in different phases of cell cycle treated with different concentrations of naringenin. Data is representative of three independent experiments.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4199682&req=5

pone-0110003-g006: Effect of naringenin on different phases of cell cycle.A431 cells were treated with different concentrations of naringenin (100 µM−300 µM) for 24 h, stained with propidium iodide and measured by flow cytometry (A) Representative photomicrograph showing the apoptosis and phase distribution of cell population (B) Quantitative distribution of percentage of A431 cells in different phases of cell cycle treated with different concentrations of naringenin. Data is representative of three independent experiments.

Mentions: Cell cycle analysis with cellular DNA content was performed by flow cytometry. Apoptotic cells were estimated by calculating the number of sub-diploid cells in the cell cycle histogram. When cells were exposed to 100 µM of naringenin, apoptotic cells reported were 6.24% as compared to control. However, at 300 µM and 500 µM of naringenin treatment, apoptotic cells were markedly increased to approximately 14.39% and 26.32% respectively. Analysis of cell cycle revealed that the cells in G0/G1 phase were observed to be 35.86%. However, 100 µM of naringenin arrest the hypodiploid cells approximately 53.75% in G0/G1 phase while 300 µM and 500 µM of naringenin arrest the cells 54.03% and 48.20% as compared to control (Fig. 6A and 6B).


Induction of apoptosis and antiproliferative activity of naringenin in human epidermoid carcinoma cell through ROS generation and cell cycle arrest.

Ahamad MS, Siddiqui S, Jafri A, Ahmad S, Afzal M, Arshad M - PLoS ONE (2014)

Effect of naringenin on different phases of cell cycle.A431 cells were treated with different concentrations of naringenin (100 µM−300 µM) for 24 h, stained with propidium iodide and measured by flow cytometry (A) Representative photomicrograph showing the apoptosis and phase distribution of cell population (B) Quantitative distribution of percentage of A431 cells in different phases of cell cycle treated with different concentrations of naringenin. Data is representative of three independent experiments.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4199682&req=5

pone-0110003-g006: Effect of naringenin on different phases of cell cycle.A431 cells were treated with different concentrations of naringenin (100 µM−300 µM) for 24 h, stained with propidium iodide and measured by flow cytometry (A) Representative photomicrograph showing the apoptosis and phase distribution of cell population (B) Quantitative distribution of percentage of A431 cells in different phases of cell cycle treated with different concentrations of naringenin. Data is representative of three independent experiments.
Mentions: Cell cycle analysis with cellular DNA content was performed by flow cytometry. Apoptotic cells were estimated by calculating the number of sub-diploid cells in the cell cycle histogram. When cells were exposed to 100 µM of naringenin, apoptotic cells reported were 6.24% as compared to control. However, at 300 µM and 500 µM of naringenin treatment, apoptotic cells were markedly increased to approximately 14.39% and 26.32% respectively. Analysis of cell cycle revealed that the cells in G0/G1 phase were observed to be 35.86%. However, 100 µM of naringenin arrest the hypodiploid cells approximately 53.75% in G0/G1 phase while 300 µM and 500 µM of naringenin arrest the cells 54.03% and 48.20% as compared to control (Fig. 6A and 6B).

Bottom Line: The search for antiproliferative agents that reduce skin carcinoma is a task of great importance.It also caused naringenin-mediated epidermoid carcinoma apoptosis by inducing mitochondrial depolarization.Cell cycle study showed that naringenin induced cell cycle arrest in G0/G1 phase of cell cycle and caspase-3 analysis revealed a dose dependent increment in caspase-3 activity which led to cell apoptosis.

View Article: PubMed Central - PubMed

Affiliation: Department of Zoology, Shibli National (PG) College, Azamgarh, Uttar Pradesh, India.

ABSTRACT
A natural predominant flavanone naringenin, especially abundant in citrus fruits, has a wide range of pharmacological activities. The search for antiproliferative agents that reduce skin carcinoma is a task of great importance. The objective of this study was to analyze the anti-proliferative and apoptotic mechanism of naringenin using MTT assay, DNA fragmentation, nuclear condensation, change in mitochondrial membrane potential, cell cycle kinetics and caspase-3 as biomarkers and to investigate the ability to induce reactive oxygen species (ROS) initiating apoptotic cascade in human epidermoid carcinoma A431 cells. Results showed that naringenin exposure significantly reduced the cell viability of A431 cells (p<0.01) with a concomitant increase in nuclear condensation and DNA fragmentation in a dose dependent manner. The intracellular ROS generation assay showed statistically significant (p<0.001) dose-related increment in ROS production for naringenin. It also caused naringenin-mediated epidermoid carcinoma apoptosis by inducing mitochondrial depolarization. Cell cycle study showed that naringenin induced cell cycle arrest in G0/G1 phase of cell cycle and caspase-3 analysis revealed a dose dependent increment in caspase-3 activity which led to cell apoptosis. This study confirms the efficacy of naringenin that lead to cell death in epidermoid carcinoma cells via inducing ROS generation, mitochondrial depolarization, nuclear condensation, DNA fragmentation, cell cycle arrest in G0/G1 phase and caspase-3 activation.

Show MeSH
Related in: MedlinePlus