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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

Effects of MEPT on the formations of acidic vesicular organelle (AVO) in HSC-4 cells. Cells were treated with vehicle or 25, 50, 100, or 150 μg/mL of MEPT for 24 h, then stained with acridine orange, and visualized by fluorescent microscopy. AVO staining at MEPT concentrations of (a) 0 μg/mL; (b) 25 μg/mL; (c) 50 μg/mL; (d) 100 μg/mL; (e) 150 μg/mL; and (f) Earle's balanced salt solution (EBSS, positive control for autophagy) (original magnification, ×400).
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fig4: Effects of MEPT on the formations of acidic vesicular organelle (AVO) in HSC-4 cells. Cells were treated with vehicle or 25, 50, 100, or 150 μg/mL of MEPT for 24 h, then stained with acridine orange, and visualized by fluorescent microscopy. AVO staining at MEPT concentrations of (a) 0 μg/mL; (b) 25 μg/mL; (c) 50 μg/mL; (d) 100 μg/mL; (e) 150 μg/mL; and (f) Earle's balanced salt solution (EBSS, positive control for autophagy) (original magnification, ×400).

Mentions: As shown in Figure 4, AVO formation was not detected in the vehicle-treated group or 25 μg/mL MEPT-treated cells. However, distinct punctate formation, the hallmark of autophagy induction, was observed in HSC-4 cells treated with >50 μg/mL MEPT (Figure 4).


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)

Effects of MEPT on the formations of acidic vesicular organelle (AVO) in HSC-4 cells. Cells were treated with vehicle or 25, 50, 100, or 150 μg/mL of MEPT for 24 h, then stained with acridine orange, and visualized by fluorescent microscopy. AVO staining at MEPT concentrations of (a) 0 μg/mL; (b) 25 μg/mL; (c) 50 μg/mL; (d) 100 μg/mL; (e) 150 μg/mL; and (f) Earle's balanced salt solution (EBSS, positive control for autophagy) (original magnification, ×400).
© Copyright Policy - open-access
Related In: Results  -  Collection

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fig4: Effects of MEPT on the formations of acidic vesicular organelle (AVO) in HSC-4 cells. Cells were treated with vehicle or 25, 50, 100, or 150 μg/mL of MEPT for 24 h, then stained with acridine orange, and visualized by fluorescent microscopy. AVO staining at MEPT concentrations of (a) 0 μg/mL; (b) 25 μg/mL; (c) 50 μg/mL; (d) 100 μg/mL; (e) 150 μg/mL; and (f) Earle's balanced salt solution (EBSS, positive control for autophagy) (original magnification, ×400).
Mentions: As shown in Figure 4, AVO formation was not detected in the vehicle-treated group or 25 μg/mL MEPT-treated cells. However, distinct punctate formation, the hallmark of autophagy induction, was observed in HSC-4 cells treated with >50 μg/mL MEPT (Figure 4).

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