• Journal of Ginseng Research
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• 제41권3호
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• pp.247-256
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• 2017

Background: KG-135, a standardized formulation enriched with Rk1, Rg3, and Rg5 ginsenosides, has been shown to inhibit various types of cancer cells; however, the underlying mechanisms are not fully understood. In this study, we explored its effects in A549 human lung cancer cells to investigate the induction of Forkhead Class box O3a (FOXO3a) and autophagy. Methods: Cell viability was determined by sulforhodamine B staining. Apoptosis and cell cycle distribution were analyzed using flow cytometry. The changes of protein levels were determined using Western blot analysis. Autophagy induction was monitored by the formation of acidic vesicular organelles stained with acridine orange. Results: KG-135 effectively arrested the cells in G1 phase with limited apoptosis. Accordingly, a decrease of cyclin-dependent kinase-4, cyclin-dependent kinase-6, cyclin D1, and phospho-retinoblastoma protein, and an increase of p27 and p18 proteins were observed. Intriguingly, KG-135 increased the tumor suppressor FOXO3a and induced the accumulation of autophagy hallmark LC3-II and acidic vesicular organelles without an increase of the upstream marker Beclin-1. Unconventionally, the autophagy adaptor protein p62 (sequestosome 1) was increased rather than decreased. Blockade of autophagy by hydroxychloroquine dramatically potentiated KG-135-induced FOXO3a and its downstream (FasL) ligand accompanied by the cleavage of caspase-8. Meanwhile, the decrease of Bcl-2 and survivin, as well as the cleavage of caspase-9, were also drastically enhanced, resulting in massive apoptosis. Conclusion: Besides arresting the cells in G1 phase, KG-135 increased FOXO3a and induced an unconventional autophagy in A549 cells. Both the KG-135-activated extrinsic FOXO3a/FasL/caspase-8 and intrinsic caspase-9 apoptotic pathways were potentiated by blockade of autophagy. Combination of KG-135 and autophagy inhibitor may be a novel strategy as an integrative treatment for cancers.

• 생명과학회지
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• 제23권6호
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• pp.812-824
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• 2013

Reserpine은 항고혈압제로서 알츠하이머질병의 증상을 나타내는 Caenorhabditis elegans에서 세포독성을 감소시켜 마비를 억제하고 수명을 연장시키는 것으로 알려져 있다. 본 연구에서는 이러한 reserpine의 효능을 포유동물에서 확인하기 위하여, 알츠하이머질병의 병리적 특성과 연관된 주요인자의 변화를 30일 동안 reserpine을 투여한 Tg2576 마우스에서 관찰하였다. 그 결과, 공격행동(aggressive behavior)은 vehicle 투여 그룹에 비하여 reserpine 투여 그룹에서 유의적으로 감소하였으나 사회적 접촉(social contact)은 유의적인 변화가 없었다. 뇌의 해마부분에서 알츠하이머질병의 원인 중 하나인 $A{\beta}$-42의 축적은 reserpine 투여 그룹에서 유의적으로 감소하였고, $A{\beta}$-42의 농도도 대조군에 비하여 reserpine 투여 그룹에서 감소하였다. 더불어, ${\gamma}$-secretase의 구성단백질 중에서 PS-2, Pen-2, APH-1의 발현은 대조군에 비하여 reserpine 투여군에서 유의적으로 감소하였으나 NCT 발현은 변화가 없었다. 혈청에서 NGF의 농도는 Tg2576 마우스에서 감소하였다가 reserpine 투여한 그룹에서 유의적으로 증가하였으며, high affinity receptor의 신호전달과정에 포함된 단백질 중에서 reserpine 투여 그룹은 TrkA의 인산화가 증가하고 ERK 인산화는 감소되었다. 한편 low affinity receptor의 신호전달과정에서, $p75^{NTR}$과 Bcl-2의 발현은 vehicle 그룹에 비하여 reserpine 투여 그룹에서 유의적으로 발현이 증가하였으나 RhoA의 발현은 reserpine 투여 그룹에서 감소하였다. 따라서 이러한 결과는 reserpine은 포유동물 치매모델인 Tg2576 마우스에서 행동학적 변화, $A{\beta}$-42의 축적, NGF의 농도, NGF신호전달의 변화 등을 유도하며, 향후 치매치료제로서 가능성을 제시하고 있다.

• 한국식품영양과학회지
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• 제42권11호
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• pp.1717-1726
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• 2013

본 연구에서는 김치 유산균 starter의 사용이 김치의 품질특성과 건강기능성에 미치는 영향을 확인하고자 single starter 김치(Lb. plantarum pnuK, Lb. plantarum 3099K, Leu. mesenteroides pnuK), mixed starter 김치(Lb. plantarum pnu/Leu. mesenteroides pnuK, Lb. plantarum 3099/Leu. mesenteroides pnuK), 자연 발효 김치(NK)를 제조하여 $15^{\circ}C$에서 6일간 발효하며 실험하였다. pH 및 산도는 발효 초기에는 차이가 없었으나 발효 2일부터 starter 접종 김치에서 빠른 변화를 보였다. 총호기성 세균과 Lactobacillus sp.는 발효가 진행될수록 비슷하게 증가하였으나 Leuconostoc sp.는 Leu. mesenteroides를 starter로 접종한 김치에서 높게 나타났다. 관능검사 결과 starter를 첨가한 김치에서 관능적 만족도가 높게 나타났으며, 혼합 starter 김치에서 만족도가 더 높았다. DPPH 및 hydroxyl radical 소거능 측정 결과, starter를 첨가한 김치에서 높은 항산화 효과를 보였다. MTT assay 결과, HCT-116 및 HT-29 세포에서 NK는 각각 63.4, 51.9%를 나타내었고 Lb. plantarum pnuK는 각각 77.1, 68.8%의 암세포 성장 저해율을 보였다. 따라서 starter 첨가 시 항산화 및 항암효과가 증가되며, 단일 starter 김치가 혼합 starter 김치보다 높은 기능성을 가지는 것으로 나타났다. 그중 새로 분리 동정된 Lb. plantarum pnu를 이용한 김치에서의 높은 대장암 세포 성장억제 효과를 확인하였다. Apoptosis 유도에 관련된 Bcl-2 및 Bax, 종양 억제에 관련된 p53 및 p21, 염증에 관련된 iNOS 및 COX-2 유전자 발현은 starter 첨가 김치에서 자연발효 김치보다 모두 apoptosis 유도 작용과 염증 및 종양을 억제하는 효과가 높게 나타났다. 가장 높은 효과를 보인 것은 Lb. plantarum pnuK이었으며 단일 starter 김치에서 혼합 starter 김치보다 효과가 높았다. 이상의 결과로 starter를 이용해 관능적 품질이 우수하고 기능성이 증진된 김치를 제조할 수 있을 것으로 기대된다.

• 생명과학회지
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• 제29권11호
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• pp.1227-1234
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• 2019

매실은 한국, 일본 및 중국 등지에서 섭취하여온 과실로서 유기산이 풍부하며, 다량의 무기질 및 폴리페놀 성분을 함유한 것으로 알려져 있다. 이러한 매실을 이용하여 예로부터 절임이나 청 등으로 가공하여 섭취할 뿐만 아니라 구토, 해독 및 해열에 효능이 있는 한약재로서 사용해왔다. 본 연구에서는 매실을 이용하여 분획물을 제조하고 각각의 인체 대장암세포에 대한 성장 억제효과를 확인하였으며, colony 형성능, 형태학적 관찰, apoptotic body 형성, 단백질 변화 및 환경호르몬에 대한 방어 효과를 평가하였다. 매실 분획물의 인체 대장암세포 HT-29에 대한 증식 억제효과는 매실 메탄올 분획물(maesil methanol fraction, MMF) 및 매실 부탄올 분획물(maesil, butanol fraction, MBF)이 가장 뛰어난 효능을 나타내었다. MMF 및 MBF의 처리에 따라 colony 형성능 또한 농도의존적으로 감소하는 것으로 나타났으며 형태학적 변화 또한 확인할 수 있었다. 핵의 형태학적 변화를 관찰한 결과 $125{\mu}g/ml$의 MMF 및 MBF 처리에 따라 apoptotic body의 형성 및 핵 응축을 관찰하였다. MMF 및 MBF의 처리에 따른 인체 대장암세포의 성장 억제효과가 apoptosis 와 관련이 있는지 단백질의 변화를 확인한 결과, pro-apoptotic 단백질은 Bax, caspase-3, PARP의 발현이 농도의존적으로 증가하는 경향을 나타내었으며, anti-apoptotic 단백질인 Bcl-2의 발현은 감소하는 경향을 나타냈다. 또한, 환경호르몬의 일종은 bisphenol A (BPA)에 의해 성장이 증식된 HT-29세포는 MMF의 처리에 따라 유의적인 성장억제 효과를 갖는 것을 확인할 수 있었다. 따라서 본 연구를 통해 얻은 결과는 매실 분획물을 이용한 기능성 식품 소재화에 대한 기초 자료로 활용될 수 있을 것으로 판단된다.

• Biomolecules & Therapeutics
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• 제17권4호
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• pp.403-411
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• 2009

Thiacremonone, the main component isolated from heated garlic (Allium sativum L.), is interested for using as a cancer preventive or therapeutic agent since garlic has been known to be useful plant in the treatment of cancers. Nuclear factor kappaB (NF-${\kappa}B$) is constitutively activated in the prostate cancer and activation of NF-${\kappa}B$ is implicated in drug resistance in cancer cells. Docetaxel, a semisynthetic analog of paclitaxel, is an antineoplastic drug widely used for advanced various cancer. In previous studies, we found that thiacremonone inhibited activation of NF-${\kappa}B$ in cancer cells and marcrophages. In the present study, we investigated whether thiacremonone could increase susceptibility of prostate cancer cells (PC-3 and DU145) to docetaxel via inactivation of NF-${\kappa}B$. We found that the combination treatment of thiacremonone (50 ${\mu}g$/ml) with docetaxel (5 nM) was more effective in the inhibition of prostate cancer cell growth and induction of apoptosis accompanied with the significant inhibition of NF-${\kappa}B$ activity than those by the treatment of thiacremonone or docetaxel alone. It was also found that NF-${\kappa}B$ target gene expression of Bax, caspase-3 and caspase-9 was much more significantly enhanced, but the expression of Bcl-2 was also much more significantly inhibited by the combination treatment. These results indicate that thiacremonone inhibits NF-${\kappa}B$, and enhances the susceptibility of prostate cancer cells to docetaxel. Thus, thiacremonone could be useful as an adjuvant anti-cancer agent.

• 동의생리병리학회지
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• 제18권5호
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• pp.1449-1455
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• 2004

Bufonis venenum (dried toad venom; Chinese name, Chan su) is a traditional Chinese medicine obtained from the skin venom gland of the toad. It has long been used in treating arrhythmia and other heart diseases in China and other Asian countries. Additionally, Bufonis venenum has been reported to selectively inhibit the growth of various lines of human cancer cells. In the present study, it was examined the effects of extract of Bufonis venenum (EBV) on the growth of human bladder carcinoma cell line T24 in order to investigate the anti-proliferative mechanism and induction of apoptosis by EBV. Treatment of T24 cells to EBV resulted in the growth inhibition, morphological change and induction of apoptotic cell death in a dose-dependent manner. Flow cytometric analysis revealed that EBV treatment caused G2/M phase arrest of the cell cycle and down-regulation of cyclin A, cyclin B1 and Cdc2, which was associated with a marked up-regulation of cyclin-dependent kinases (Cdks) inhibitor p21 (WAF1/CIP1) in a p53-independent manner. The Cdc25C expression was also significantly inhibited by EBV treatment, however Wee1 kinase expression was not affected. The induction of apoptotic cell death by EBV was connected with down-regulation of anti-apoptotic Bcl-XS/L expression without alteration pro-apoptotic Bax expression. Taken together, these findings suggest that EBV may be a potential chemotherapeutic agent for the control of human bladder carcinorma cells and further studies will be needed to identify the active compounds that confer the anti-cancer activity of EBV.

• 동의생리병리학회지
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• 제21권5호
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• pp.1226-1232
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• 2007

Phellinus linteus is a well-known Oriental medicinal fungus that has various biological activities, including immunomodulatory and anti-tumor activities, the mechanisms of which are poorly understood. In the present study, we investigated the anti-proliferative activity of the methanol extract of P. linteus-Barley corn (MEPLB) in human lekemic U937 cells. It was found that exposure of U937 cells to MEPLB resulted morphological change and growth inhibition in a dose-dependent manner as measured by trypan blue count and MTT assay. Upon treatment with MEPLB, U937 cells developed many of the hallmark features of apoptosis, including condensation of chromatin and an increase in the sub-G1 population suggesting that the anti-proliferative effect of MEPLB is associated with the induction of apoptosis. The anti-proliferative and apoptotic effects of MEPLB were connected with a marked induction of the pro-apoptotic Bax and cyclin-dependent kinase (Cdk) inhibitor p21 in a p53-independent manner. Additionally, MEPLB treatment significantly induced the caspase-3 activity in U937 cells. Taken together, the present results suggest that apoptotic signals evoked by MEPLB in human leukemic U937 cells may converge caspase-3 activation through an up-regulation of Bax rather than a down-regulation of Bcl-2 or Bel-xL.

• Journal of Nutrition and Health
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• 제53권4호
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• pp.369-380
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• 2020

본 연구에서는 일반식이로의 식이 제한이 고지방식이로 촉진된 유방암의 성장과 전이에 미치는 영향과 그 기전을 알아보았다. 8주간 고지방식이를 섭취시킨 후 유방암을 유방 조직에 이식하거나 꼬리 정맥으로 통해 전이시켰으며, 암 생성 및 전이 후 HFC군은 고지방식이를, DR군은 일반식이로 전환시켜 사육하였다. 본 연구 결과, 원발성 유방암의 경우, HFC군과 DR군의 체중에는 차이가 없었으나 간과 비장, 신장 주변의 지방세포의 무게에서 유의한 차이를 보였다. DR군이 HFC군에 비해 유방암의 개시를 지연시켰으며, 유방암의 성장과 무게를 유의하게 감소시켰다. 일반식이로의 전환에 의한 유방암 성장 억제는 세포 분열 억제와 세포사멸을 조절하는 인자의 발현 감소에 의한 caspase-3의 활성 촉진에 의한 것으로 나타났다. 또한, 식이 제한은 폐로 전이된 유방암의 수를 유의미하게 감소시켰다. 본 실험 결과, 일반식이로의 제한은 고지방식이로 촉진된 유방암의 성장과 전이를 억제하는 효과를 나타내며, 이는 유방암 세포의 성장 억제와 사멸 유도에 의한 것으로 나타났다. 따라서, 유방암의 성장이나전이를 억제하기 위해서는 총 섭취 열량을 조절하고, 지방의 섭취량을 줄이는 균형 잡힌 식이를 섭취하도록 해야 할 것으로 사료된다.

• Biomolecules & Therapeutics
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• 제24권6호
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• pp.623-629
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• 2016

(1S,2S,3E,7E,11E)-3,7,11,15-cembratetraen-17,2-olide (LS-1), a marine cembrenolide diterpene, has anticancer activity against colon cancer cells such as HT-29, SNU-C5/5-FU (fluorouracil-resistant SNU-C5) and SNU-C5. However, the action mechanism of LS-1 on SNU-C5 human colon cancer cells has not been fully elucidated. In this study, we investigated whether the anticancer effect of LS-1could result from apoptosis via the modulation of $Wnt/{\beta}$-catenin and the TGF-${\beta}$ pathways. When treated with the LS-1, we could observe the apoptotic characteristics such as apoptotic bodies and the increase of sub-G1 hypodiploid cell population, increase of Bax level, decrease of Bcl-2 expression, cleavage of procaspase-3 and cleavage of poly (ADP-ribose) polymerase in SNU-C5 cells. Furthermore, the apoptosis induction of SNU-C5 cells upon LS-1 treatment was also accompanied by the down-regulation of $Wnt/{\beta}$-catenin signaling pathway via the decrease of GSK-$3{\beta}$ phosphorylation followed by the decrease of ${\beta}$-catenin level. In addition, the LS-1 induced the activation of TGF-${\beta}$ signaling pathway with the decrease of carcinoembryonic antigen which leads to decrease of c-Myc, an oncoprotein. These data suggest that the LS-1 could induce the apoptosis via the down-regulation of $Wnt/{\beta}$-catenin pathway and the activation of TGF-${\beta}$ pathway in SNU-C5 human colon cancer cells. The results support that the LS-1 might have potential for the treatment of human colon cancer.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제35권3호
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• pp.143-154
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• 2013

Purpose: This study was to find efficacy of integrin alpha2 (${\alpha}_2$) and epidermal growth factor receptor (EGFR) as tumor marker of oral squamous cell carcinoma (SCC) and clarify the selective cell death effect of anti-integrin ${\alpha}_2$ and anti-EGFR on SCC cells, additionally testify conjugated gold nanoparticles (GNP) with air plasma for selective cell death of oral SCC. Methods: Expression of integrin ${\alpha}_2$, EGFR on human SCC cells (SCC25) were examined by western blot. SCC25 cells were treated with anti-integrin ${\alpha}_2$, anti-EGFR and analysed by Hemacolor staining, immunoflorescence staining, FACS flow cytometry. Conjugated GNP with integrin ${\alpha}_2$, EGFR antibody were treated by air plasma on SCC cells. Results: Integrin ${\alpha}_2$ and EGFR were over-expressed on SCC25 cells than normal lung WI-38 cells. The cell viability rate of SCC25 cells treated with anti-integrin ${\alpha}_2$, anti-EGFR was lower than WI-38 cells. The concentration changes of nucleus, releasing cytochrome c and apoptosis inducing factor (AIF) from mitochondria to cytosol were observed. The changes of proteins related with apoptosis were observed. Increase of bax, bcl-xL, activation of caspase-3, -7, -9, and fragmentation of PARP, DFF45 and decrease of lamin A/C in SCC25 cells were observed. In FACS, increase of sub-$G_1$ and S phase was observed. Cell cycle related proteins, Such as cyclin D1, cyclin dependent kinase (CDK) 4, cyclin A, cyclin E, CDK 2, p27 were decreased. After SCC25 cells treated with conjugatged GNP-Integrin ${\alpha}_2$, GNP-EGFR, additionally air plasma, the cell death rate was significantly increased. Conclusion: Integrin ${\alpha}_2$, EGFR were over-expressed in oral SCC cells. Anti-integrin ${\alpha}_2$, anti-EGFR in SCC25 cells induced apoptosis selectively. When GNP-anti integrin ${\alpha}_2$, GNP-anti EGFR were treated with air plasma on SCC25 cells, cancer cells were died more selectively. GNP-anti integrin ${\alpha}_2$, GNP-anti EGFR with air plasma could be treatment choice of oral SCC.