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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 viabilities of HSC-4 cells pretreated with 3-MA. HSC-4 cells were pretreated with or without 3-MA for 1 h and then 25, 50, or 100 μg/mL of MEPT for 24 h, after which cell viability was analyzed using an MTT assay. Results are expressed as the means ± standard deviations. ∗∗∗P < 0.001 versus the same amount of MEPT-treated cells.
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fig6: Effects of MEPT on the viabilities of HSC-4 cells pretreated with 3-MA. HSC-4 cells were pretreated with or without 3-MA for 1 h and then 25, 50, or 100 μg/mL of MEPT for 24 h, after which cell viability was analyzed using an MTT assay. Results are expressed as the means ± standard deviations. ∗∗∗P < 0.001 versus the same amount of MEPT-treated cells.

Mentions: To determine the type of MEPT-induced autophagy, we conducted an MTT assay of HSC-4 cells pretreated with 3-MA (a classical inhibitor of autophagy) for 1 h prior to MEPT treatment. The results revealed that the cell viability of the 3-MA pretreated and MEPT-treated cells was significantly greater than that of only MEPT-treated cells (Figure 6).


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 viabilities of HSC-4 cells pretreated with 3-MA. HSC-4 cells were pretreated with or without 3-MA for 1 h and then 25, 50, or 100 μg/mL of MEPT for 24 h, after which cell viability was analyzed using an MTT assay. Results are expressed as the means ± standard deviations. ∗∗∗P < 0.001 versus the same amount of MEPT-treated cells.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig6: Effects of MEPT on the viabilities of HSC-4 cells pretreated with 3-MA. HSC-4 cells were pretreated with or without 3-MA for 1 h and then 25, 50, or 100 μg/mL of MEPT for 24 h, after which cell viability was analyzed using an MTT assay. Results are expressed as the means ± standard deviations. ∗∗∗P < 0.001 versus the same amount of MEPT-treated cells.
Mentions: To determine the type of MEPT-induced autophagy, we conducted an MTT assay of HSC-4 cells pretreated with 3-MA (a classical inhibitor of autophagy) for 1 h prior to MEPT treatment. The results revealed that the cell viability of the 3-MA pretreated and MEPT-treated cells was significantly greater than that of only MEPT-treated cells (Figure 6).

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