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KMID : 0895820070170010027
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Journal of Oriental Rehabilitation Medicine
2007 Volume.17 No. 1 p.27 ~ p.41
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Effect of Gamihangbak-tang(Jiaweikangbai-tang) on Apoptosis of HL-60 Cells
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Chung Sang-Pil
Kwon Young-Dal
Song Yung-Sun
Lee Su-Kyung
Kim Kyeong-Su
Park Seong-Heak
Jung Moon-Jae
Song Jung-Sup
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Abstract
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Objectives: To investigate anti-leukemic effect of Gamihangbak-tang(Jiaweikangbai-tang) using human promyelocytic leukemic HL-60 cells.
Methods: Cell viability was determined using MTT assay. Morphological change was observed under a fluorescence microscope through DAPI-Methol staining. DNA fragmentation was analyzed by agarose gel electrophoresis. Caspase activity was measured by using a fluorogenic caspase substrate. Cleavage of caspase-3, PARP and Bid, expression of Bcl-2 and Bax, release of cytochrome C, and activation of MAPKs (ERK, JNK, p38) were determined by Western blot analysis.
Results: Gamihangbak-tang(Jiaweikangbai-tang) treated HL-60 cells increased time-dependently DNA fragments and apoptotic bodies. Activities of caspase-3, 8, 9 increased on 30min after Gamihangbak-tang(Jiaweikangbai-tang) treatment and greatly increased after 3h treatment. PARP cleavage also increased 89 kDa PARP fragment on 3hr after Gamihangbak-tang(Jiaweikangbai-tang) treatment. Expression of Bcl-2 and Bax in Gamihangbak-tang (Jiaweikangbai-tang) treated HL-60 cells was not changed, but caspase-8-dependent Bid cleavage was observed. Gamihangbak-tang Jiaweikangbaitang) also induced cytochrome C release from mitochondria into the cytosol. Activation of ERK, p38, and JNK increased in apoptosis induced by Gamihangbak-tang(Jiaweikangbai-tang) in HL-60 cells, but only ERK activation was concerned in Gamihangbak-tang(Jiaweikangbai-tang)-induced apoptosis.
Conclusions: Apoptosis induced by Gamihangbak-tang(Jiaweikangbai-tang) in HL-60 cells induce caspase-3 activation through Bid cleavage by caspase-8, caused cytochrome C release from mitochondria, and ERK activation.
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KEYWORD
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HL-60, Gamihangbak-tang(Jiaweikangbai-tang), Apoptosis, ERK, p38, JNK
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