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Autophagic Cell Death by Poncirus trifoliata Rafin., a Traditional Oriental Medicine, in Human Oral Cancer HSC-4 Cells.

Han HY, Park BS, Lee GS, Jeong SH, Kim H, Ryu MH - Evid Based Complement Alternat Med (2015)

Bottom Line: The methanol extract of P. trifoliata (MEPT) significantly decreased the proliferation of HSC-4 cells (inhibitory concentration (IC)50 = 142.7 μg/mL) in a dose-dependent manner.While there were no significant changes observed upon cell cycle analysis and ANNEXIN V and 7-AAD double staining in the MEPT-treated groups, the intensity of acidic vesicular organelle (AVO) staining and microtubule-associated protein 1 light chain (LC) 3-II protein expression increased in response to MEPT treatment.Taken together, our results indicate that MEPT is a potent autophagy agonist in oral cancer cells with antitumor therapeutic potential that acts through the mitogen-activated protein kinase (MAPK) pathway.

View Article: PubMed Central - PubMed

Affiliation: Department of Oral Pathology, School of Dentistry, Institute of Translational Dental Sciences, Pusan National University, Yangsan, Republic of Korea.

ABSTRACT
Poncirus trifoliata Rafin. has long been used as anti-inflammatory and antiallergic agent to treat gastrointestinal disorders and pulmonary diseases such as indigestion, constipation, chest fullness, chest pain, bronchitis, and sputum in Korea. P. trifoliata extract has recently been reported to possess anticancer properties; however, its mechanisms of action remain unclear. In this study, its antiproliferative effects and possible mechanisms were investigated in HSC-4 cells. The methanol extract of P. trifoliata (MEPT) significantly decreased the proliferation of HSC-4 cells (inhibitory concentration (IC)50 = 142.7 μg/mL) in a dose-dependent manner. While there were no significant changes observed upon cell cycle analysis and ANNEXIN V and 7-AAD double staining in the MEPT-treated groups, the intensity of acidic vesicular organelle (AVO) staining and microtubule-associated protein 1 light chain (LC) 3-II protein expression increased in response to MEPT treatment. Furthermore, 3-methyladenine (3-MA, autophagy inhibitor) effectively blocked the MEPT-induced cytotoxicity of HSC-4 cells and triggered the activation of p38 and extracellular signal-regulated kinases (ERK) proteins. Taken together, our results indicate that MEPT is a potent autophagy agonist in oral cancer cells with antitumor therapeutic potential that acts through the mitogen-activated protein kinase (MAPK) pathway.

No MeSH data available.


Related in: MedlinePlus

The expression of LC3 and Atg proteins in MEPT-treated HSC-4 cells. Western blotting was performed for LC3-I and LC3-II, Atg4B, and Atg5-12 conjugate protein after administering MEPT at 0, 25, 50, or 100 μg/mL for 24 h or 10 μM TFP (positive control for autophagy) for 24 h.
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fig5: The expression of LC3 and Atg proteins in MEPT-treated HSC-4 cells. Western blotting was performed for LC3-I and LC3-II, Atg4B, and Atg5-12 conjugate protein after administering MEPT at 0, 25, 50, or 100 μg/mL for 24 h or 10 μM TFP (positive control for autophagy) for 24 h.

Mentions: As shown in Figure 5, MEPT treatment converted LC3-I into LC3-II isoform and activated Atg4B protein. However, the level of Atg5-12 conjugate was not affected by MEPT (Figure 5).


Autophagic Cell Death by Poncirus trifoliata Rafin., a Traditional Oriental Medicine, in Human Oral Cancer HSC-4 Cells.

Han HY, Park BS, Lee GS, Jeong SH, Kim H, Ryu MH - Evid Based Complement Alternat Med (2015)

The expression of LC3 and Atg proteins in MEPT-treated HSC-4 cells. Western blotting was performed for LC3-I and LC3-II, Atg4B, and Atg5-12 conjugate protein after administering MEPT at 0, 25, 50, or 100 μg/mL for 24 h or 10 μM TFP (positive control for autophagy) for 24 h.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4499625&req=5

fig5: The expression of LC3 and Atg proteins in MEPT-treated HSC-4 cells. Western blotting was performed for LC3-I and LC3-II, Atg4B, and Atg5-12 conjugate protein after administering MEPT at 0, 25, 50, or 100 μg/mL for 24 h or 10 μM TFP (positive control for autophagy) for 24 h.
Mentions: As shown in Figure 5, MEPT treatment converted LC3-I into LC3-II isoform and activated Atg4B protein. However, the level of Atg5-12 conjugate was not affected by MEPT (Figure 5).

Bottom Line: The methanol extract of P. trifoliata (MEPT) significantly decreased the proliferation of HSC-4 cells (inhibitory concentration (IC)50 = 142.7 μg/mL) in a dose-dependent manner.While there were no significant changes observed upon cell cycle analysis and ANNEXIN V and 7-AAD double staining in the MEPT-treated groups, the intensity of acidic vesicular organelle (AVO) staining and microtubule-associated protein 1 light chain (LC) 3-II protein expression increased in response to MEPT treatment.Taken together, our results indicate that MEPT is a potent autophagy agonist in oral cancer cells with antitumor therapeutic potential that acts through the mitogen-activated protein kinase (MAPK) pathway.

View Article: PubMed Central - PubMed

Affiliation: Department of Oral Pathology, School of Dentistry, Institute of Translational Dental Sciences, Pusan National University, Yangsan, Republic of Korea.

ABSTRACT
Poncirus trifoliata Rafin. has long been used as anti-inflammatory and antiallergic agent to treat gastrointestinal disorders and pulmonary diseases such as indigestion, constipation, chest fullness, chest pain, bronchitis, and sputum in Korea. P. trifoliata extract has recently been reported to possess anticancer properties; however, its mechanisms of action remain unclear. In this study, its antiproliferative effects and possible mechanisms were investigated in HSC-4 cells. The methanol extract of P. trifoliata (MEPT) significantly decreased the proliferation of HSC-4 cells (inhibitory concentration (IC)50 = 142.7 μg/mL) in a dose-dependent manner. While there were no significant changes observed upon cell cycle analysis and ANNEXIN V and 7-AAD double staining in the MEPT-treated groups, the intensity of acidic vesicular organelle (AVO) staining and microtubule-associated protein 1 light chain (LC) 3-II protein expression increased in response to MEPT treatment. Furthermore, 3-methyladenine (3-MA, autophagy inhibitor) effectively blocked the MEPT-induced cytotoxicity of HSC-4 cells and triggered the activation of p38 and extracellular signal-regulated kinases (ERK) proteins. Taken together, our results indicate that MEPT is a potent autophagy agonist in oral cancer cells with antitumor therapeutic potential that acts through the mitogen-activated protein kinase (MAPK) pathway.

No MeSH data available.


Related in: MedlinePlus