• 동의생리병리학회지
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• 제20권2호
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• pp.345-351
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• 2006

We have examined the induction of HL-60 cell differentiation by treatment of the water extract of Rhizoma Zingiberis Siccatum, which is a Korean traditional herbal medicine. It was observed that HL-60 cell proliferation was dose-dependently inhibited by treatment with various dose of Rhizoma Zingiberis Siccatum extract. The level of inhibition was identical to those of ATRA-treated cells. Rhizoma Zingiberis Siccatum extract treatment caused a significant change in NBT reduction (47%) and enhanced ATRA-induced NBT reduction. Treatment of Rhizoma Zingiberis Siccatum to HL-60 cells increased only CD11 b expression in the cells, and also increased markedly G0/G1 stage arrest. This can suggest that Rhizoma Zingiberis Siccatum induced the differentiation of HL-60 cell and enhanced ATRA-induced differentiation predominantly along the granulocytic lineage. The results present here show that Rhizoma Zingiberis Siccatum extract contains the potential effect of differentiation induction or acts as a agent enhancing the induction of differentiation.

• 동의생리병리학회지
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• 제19권1호
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• pp.234-241
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• 2005

Galla Rhois is a nest of parasitic bug, Mellaphis chinensis Bell, in Rhus chinensis Mill. Galla Rhois has been used for the therapy of diarrhea, peptic ulcer, hemauria, etc., that showed various antiinflammatory activity, and other biological properties. We studied the effect of Galla Rhois water extract(GRWE). The cytotoxic activity of GRWE in HL-60 cells was increased in a concentration-dependent manner. GRWE was cytotoxic to HL-60 cells, with $IC_50$ of $100{\mu}g/m{\ell}$. Treatment of GRWE to HL-60 cells showed the fragmentation of DNA in a concentration manner, suggesting that these cells underwent apoptosis. In addition, the flow cytometric analysis revealed GRWE concentration-dependently increased apoptotic cells with hypodiploid DNA content and arrested G1 phase of cell cycle. These results indicate that GRWE may have a possibility of potential anticancer activities. Treatment of HL-60 cells with GRWE was induced activation of caspase-3, caspase-8 and proteolytic cleavage of poly(ADP-ribose) polymerase. Also, caspase-3 was directly activated via caspase-8 activation. GRWE also caused the release of cytochrome c from mitochondria into the cytosol. GRWE-induced cytochrome c release was mediated by caspase-8-dependent cleavage of Bid and Bax translocation. These results suggest that caspase-8 mediates caspase-3 activation and cytochrome c release during GRWE-induced apoptosis in HL-60 cells.

• 동의생리병리학회지
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• 제19권6호
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• pp.1636-1639
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• 2005

To develop novel anti-leukemic medicine, we have prepared a Korean traditional medicine, named Hang-baek-Tang, which is composed of 8 kinds of anti-leukemic medicinal plants. The water extracts was examined anti-leukemic activity using the human leukemia cell line, HL-60 cells. HL-60 cells showed the growth inhibition and several apoptotic features, including DNA ladders, morphological changes, by treatment of the cells with Hang-Daek-Tang. We have observed that Hang-baek-Tang induced the activation of caspase-3, caspase-8 and caspase-9. Further molecular analysis demonstrated that Hang-baek-Tang induced cleavage of PARP and increase of hypodiploid (Sub-G1) population in flow cytometric analysis. These results indicate that Hang-baek-Tang has been considered to exert anti-leukemic activity through the procaspase-3 activation pathway.

• Asian Pacific Journal of Cancer Prevention
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• 제13권4호
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• pp.1119-1124
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• 2012

Background: The effect and possible mechanism of traditional Chinese medicine, baicalin, on the PI3K/Akt signaling pathway in drug-resistant human myeloid leukemia HL-60/ADR cells have been investigated in this current study. Methods: HL-60/ADR cells were treated by 20, 40, $80\;{\mu}mol/L$ baicalin followed by cell cycle analysis at 24h. The mRNA expression level of the apoptosis related gene, Bcl-2 and bad, were measured by RT-PCR on cells treated with $80\;{\mu}mol/L$ baicalin at 12, 24 and 48hr. Western blot was performed to detect the changes in the expression of the proteins related to HL-60/ADR cell apoptosis and the signaling pathway before and after baicalin treatment, including Bcl-2, PARP, Bad, Caspase 3, Akt, p-Akt, NF-${\kappa}B$, p-NF-${\kappa}B$, mTOR and p-mTOR. Results: Sub-G1 peak of HL-60/ADR cells appeared 24 h after $20\;{\mu}mol/L$ baicalin treatment, and the ratio increased as baicalin concentration increased. Cell cycle analysis showed 44.9% G0/G1 phase cells 24 h after baicalin treatment compared to 39.6% in the control group. Cells treated with $80\;{\mu}mol/L$ baicalin displayed a trend in decreasing of Bcl-2 mRNA expression over time. Expression level of the Bcl-2 and PARP proteins decreased significantly while that of the PARP, Caspase-3, and Bad proteins gradually increased. No significant difference in Akt expression was observed between treated and the control groups. However, the expression levels of p-Akt, NF-${\kappa}B$, p-NF-${\kappa}B$, mTOR and p-mTOR decreased significantly in a time-dependent manner. Conclusions: We conclude that baicalin may induce HL-60/ADR cell apoptosis through the PI3K/AKT signaling pathway.

• 대한방사선협회지
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• 제30권1호
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• pp.70-76
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• 2004

To evaluate the process of radiation-induced cell damage in human promyelocytic leukemia cell HL-60 cell and chinese hamster cell CHO cell lines. I irradiated HL-60 cells, CHO cell line, using the V - radiation from Cs-137 cell irradiation(Gamma cell 3000

• 한국식품영양과학회지
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• 제38권10호
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• pp.1317-1323
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• 2009

당유자 과피(GGP) 80% 에탄올 추출물을 4종의 암세포에 (피부암, 대장암, 유방암 및 혈액암) 처리하여 증식 억제 활성을 측정한 결과, 혈액암 HL60 세포에서 높은 증식 억제 활성을 보였다. 이에 CGP 추출물이 HL60 세포에 대한 apoptosis 유도에 따른 세포 증식 억제 활성을 조사하였다. Apoptosis 유도의 첫 단계인 막 투과성을 측정한 결과, confocal image와 flow cytometry에서 CGP를 처리하였을 때 탈분극 현상에 따른 막 투과성이 증가하였고 세포내 핵을 hoechst 33342를 이용하여 염색하였을 때 apoptosis가 일어났을 때 나타나는 전형적인 형태의 apoptotic body가 농도 의존적으로 증가하는 것을 확인할 수 있었으며 flow cytometry를 통하여 세포 주기를 측정하였을 때 DNA-hypodiploid 형태의 sub-G1가 CGP 농도 의존적으로 증가하는 것을 확인할 수 있었다. Apoptosis 유도 기전을 western blot으로 측정한 결과를 보면, CGP 추출물을 혈액암 HL60 세포에 처리하였을 때 Bcl family의 anti-apoptotic Bcl-2 단백질의 감소와 pro-apoptotic Bax 단백질의 증가로 인하여 하위 기전인 caspase-3가 활성화되었으며, 이 활성화로 인하여 apoptosis 유도에 직접적으로 관여하는 PARP 단백질을 활성화시키면서 apoptosis를 유도하였다. 따라서 당유자 과피는 항암과 관련되어진 기능성식품 및 소재 개발 원료로서 개발이 가능하리라고 사료된다.

• Advances in Traditional Medicine
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• 제4권3호
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• pp.171-178
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• 2004

Ginseng radix, the root of Panax ginseng C. A. Meyer (Araliaceae), is a medicinal plant used world-widely and has been reported to have various biological effects. To investigate the effects of Ginseng radix on HL-60 cell apoptosis, MTT assay, DNA fragmentation assay and flow cytometry were performed on HL-60 cells. Cells were treated with Ginseng radix at different concentrations $(10^{-4},\;10^{-3}\;and\;10^{-2};\;dilution\;rate)$. Ginseng radix significantly induced cells apoptosis with a time- and dose-dependent manner. To determine whether Ginseng radix-induced apoptosis is due to increase of tumor necrosis factor $(TNF-{\alpha})$ secretion, enzyme-linked immunosorbent assay was performed on HL-60 cells. Unexpectedly, Ginseng radix $(96\;{\pm}\;5\;pg/ml)$ significantly decreased the $TNF-{\alpha}$ secretion compared with control $(174\;{\pm}\;14\;pg/ml)$. Furthermore, Ginseng radix with $rIFN-{\gamma}$ synergistically increased nitric oxide production in mouse peritoneal macrophages. Taken together, our data indicate that Ginseng radix induce apoptosis on HL-60 cells without increase of $TNF-{\alpha}$ secretion and could be used for a supplementary remedy of cancer.

• 동의생리병리학회지
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• 제16권6호
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• pp.1190-1196
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• 2002

Selaginella Tamariscina is widely used in the traditional oriental herbal medicine for its anti-inflammatory, anti-cancer effects. The effects of aqueous extracts of Selaginella Tamariscina (ST) on the cell viability and induction of apoptotic cell death were investigated in A549, Raw 264.7, C6-glioma. Jurkat and HL-60 cells. The cell viability after treating with extract of Selaginella Tamariscina was quantified by MTT assay method. The results showed that ST decreased the cell viability in HL-60 and Jurkat cells not in A549, Raw 264.7 and C6-glioma cells. And we also observed the chromatin condensation and DNA fragmentation in HL-60 and Jurkat cells. The enzyme activity of caspase-3, tightly regulated by an apoptosis activating complex, were markedly increased in HL-60 cells treated with the ST by dose-dependent manner. In conclusion, our results suggest that the extract of Selaginella Tamariscina may induce the selective apoptotic cell death in HL-60 and Jurkat cells via activation of caspase-3.

• Biomolecules & Therapeutics
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• 제15권3호
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• pp.145-149
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• 2007

We have previously shown that 2,4,3',5'-tetramethoxystilbene (TMS), a synthetic trans-stilbene analogue acting as a potent inhibitor of human cytochrome P450 1B1, induces apoptotic cell death in human cancer cells. In the present studies, we report the mechanisms of apoptotic cell death by TMS in human promyelocytic leukemic HL-60 cells. We found that treatment of HL-60 cells with TMS suppressed the cell growth in a concentration-dependent manner with $IC_{50}$ value of about 0.8 ${\mu}M$. Immunoblot experiments revealed that DMHS-induced apoptosis was associated with cleavage of poly (ADP-ribose) polymerase. The release of cytochrome c from mitochondria into the cytosol was significantly increased in response to TMS. TMS caused activation of caspase-3 in a concentration-dependent manner and TMS-mediated caspase-3 activation was partially prevented by the caspase inhibitor, zVAD-fmk. Interestingly, we found that the cytotoxic effect of anticancer drugs such as paclitaxel, docetaxel, or etoposide was enhanced in the presence of TMS. Simultaneous treatment with TCDD also significantly increased cytotoxic effects of TMS alone or TMS and anti-cancer agents. Taken together, our present results indicated that TMS leads to apoptotic cell death in HL-60 cells through activation of caspase-3 activity and release of cytochrome c into cytosol. The ability of TMS to increase cytotoxic effect of anticancer drugs may contribute to its usefulness for cancer chemotherapy.

• 동의생리병리학회지
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• 제21권5호
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• pp.1170-1175
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• 2007

We have examined the effect of water extract of Phellinus linteus, a raw material of Korean traditional herbal medicine, on the induction of HL-60 cell differentiation. The proliferation of HL-60 cell was inhibited dose-dependently by treatment with various doses of P. linteus extract. It also caused a significant change in NBT reduction (7.5 times). The expression of CD11b and CD14 was increased in the cells treated with the extract, especially in those arrested at G0/G1 stage, which suggested that some components in P. Linteus extract induced HL-60 cell differentiation to granulocytic and monocyte lineages. Moreover, the expression levels of $p21^{WAF1/CIP}$ and $p27^{KIP}$ were up-regulated during HL-60 cell differentiation induced by P. Linteus extract. These results together suggest that P. Linteus extract contains potential HL-60 cell differentiation agents.