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KMID : 0363620190400020035
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Journal of Korean Oriental Medicine
2019 Volume.40 No. 2 p.35 ~ p.50
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Effect of Orostachys japonicus on Apoptosis and Autophagy in Human monocytic leukemia Cell line THP-1 via Inhibition of NF-κB and Phosphorylation of p38 MAPK
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Joo Seong-Hee
Jang Eun-Gyeong
Kim Young-Chul
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Abstract
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Objectives: Orostachys japonicas (O. japonicus) has been known for its anti-tumor effect. In the present study, it was investigated whether O. japonicus EtOH extracts could induce apoptosis and autophagy which are part of the main mechanism related to anti-tumor effect in THP-1 cells.
Methods: Cells were treated with various concentrations of O. japonicus EtOH extracts (0-300μg/ml) for 24, 48, and 72h. Cell viability was evaluated by MTS/PMS assay and apoptosis rate was examined by flow cytometry and ELISA assay. The mRNA expression of apoptosis-related genes (Bcl-2, Mcl-1, Survivin, Bax) and autophagy-related gene (mTOR) was evaluated using real-time PCR. The protein expression of Caspase-3, Akt, LC3Ⅱ, Beclin-1, Atg5, NF-κB, p38, ERK was evaluated using western blot analysis.
Results: O. japonicus EtOH extracts inhibited cell proliferation and apoptosis rate was increased in both flow cytometry and ELISA assay. Bcl-2, Mcl-1, Survivin (anti-apoptosis factors) mRNA expressions were decreased and Bax (pro-apoptosis factor) mRNA level was increased. mTOR mRNA expressions was decreased and LC3Ⅱ protein expressions was increased. Activation of NF-κB was decreased and phosphorylation of p38 was increased.
Conclusion: O. japonicus is regarded to inhibit cell proliferation, to induce apoptosis and to regulate autophagy-related genes in THP-1 cells via NF-κB and p38 MAPK signaling pathway. This suggests O. japonicus could be an effective herb in treating acute myeloid leukemia.
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KEYWORD
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Orostachys japonicus, THP-1, acute myeloid leukemia, apoptosis, autophagy
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