• 생명과학회지
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• 제33권12호
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• pp.978-986
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• 2023

선천면역체계에 의한 염증은 감염에 의해 매개되는 환경적 위험 요인에 대한 보호 메커니즘이며 암 발병을 포함한 다양한 인간 질병의 발병 원인이기도 하다. 지의류는 다양한 질병을 치료할 수 있는 가능성을 지닌 다양한 생체 활성분자를 가지고 있다는 측면에서 점점 더 주목받고 있다. 이끼류의 2차 대사산물이 지닌 항산화, 항염증 그리고 항암 활성에 대해서 널리 보고되었지만 아직까지 구체적인 메커니즘은 밝혀지지 않았다. 본 연구에서는 남극 지의류 Usnea aurantiaco-atra의 메탄올 추출물에 대한 항염증 및 항암 활성의 분자적 메커니즘을 조사하고자 하였다. 본 연구결과에 의하면 메탄올 추출물은 COX-2, IL-6, iNOS, TNF-α 및 NO 생성과 같은 주요 염증 지표들에 대해 농도 의존적으로 조절함으로써 항염증 활성을 나타냈다. 또한, 추출물이 농도 의존적으로 HCT116 결장암 세포에 대해 세포독성 활성을 가지며, caspase-3 활성화에 의한 세포사멸 유도를 통해 암세포의 증식을 현저히 감소시키는 것을 관찰했다. 이 연구에서 남극 이끼류인 Usnea aurantiaco-atra의 메탄올 추출물이 항염증과 항암 활성을 갖는다는 사실을 처음으로 보였으며 이러한 결과는 염증과 암 사이의 연관성을 뒷받침하는 분자 메커니즘에 대한 새로운 통찰력을 보여준다.

• 한국축산식품학회지
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• 제38권3호
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• pp.615-628
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• 2018

Preserved egg, a kind of alkaline-fermented food, is a traditional egg product in China. Here, we investigated the nutritional functions of preserved eggs by in vivo and in vitro experiments. The results of in vivo studies showed that the levels of triglycerides (TG), total cholesterol (TCHO) and low-density lipoprotein cholesterol/high density lipoprotein cholesterol (LDL-C/HDL-C) were significantly decreased (p<0.05) in the liver of rats treated with preserved eggs. Meanwhile, the levels of two important cancer markers, interleukin-6 (IL-6) and tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$), were also significantly decreased (p<0.05) in treated rats. In vitro studies were performed on Caco-2 cells, a human epithelial colorectal adenocarcinoma cell line. It demonstrated that the gastrointestinal (GI) digests of preserved eggs significantly accelerated (p<0.05) the apoptosis by upregulating caspase-3 in the Caco-2 cells. Besides, after treated with preserved eggs, the half maximal inhibitory concentration (IC50) of preserved eggs digests to Caco-2 cells was 5.75 mg/mL, indicating the significant inhibition of cell proliferation provided by preserved eggs (p<0.05). The results shown in this study demonstrated that preserved eggs may be a novel functional food involved with antilipemic, anti-inflammatory activity as well as the effect on accelarating the apoptosis of Caco-2 cells.

• 동의생리병리학회지
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• 제22권2호
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• pp.290-295
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• 2008

The cytotoxic characteristics of Vitsin A isolated from Vitis amurensis L. were examined in human colorectal, breast, uterine and renal cancer cells. Vitsin A showed good cytotoxicity against various cancer cells with $IC_{50}$ of $1\;{\sim}\;30\;{\mu}M$. Among them, Vitisin A exhibited strongest cytotoxic effect against MES-SA cells with $IC_{50}$ of 1.11 ${\mu}M$ by SRB assay. To verify whether the cytotoxicity of Vitisin A may be associated with apoptosis, TdT-mediated-dUTP Nick-End Labeling (TUNEL) assay and cell cycle analysis were performed in MES-SA cells. Apoptotic bodies were observed in Vitisin A treated MES-SA cells by TUNEL assay. Also, Vitisin A effectively increased the portion of $sub-G_1$ DNA content by flow cytometric analysis. Taken together, these findings suggest that the cytotoxicity of Vitisin A against MES-SA cells is chiefly mediated by apoptosis.

• 고려인삼학회:학술대회논문집
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• 고려인삼학회 1998년도 Advances in Ginseng Research - Proceedings of the 7th International Symposium on Ginseng -
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• pp.270-280
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• 1998

Ginseng is one of the most widely used medicinal plants, particularly in East Asian countries. Certain fractions or purified ingredients of ginseng have been shown to exert inhibitory effects on growth of cancer cells in culture or on tumorigenesis in experimental animals. Moreover, a recent epidemiologic study reveals that ginseng intake is associated with a reduced risk for environmentally related cancers such as esophageal, gastric, colorectal, and pulmonary tumors. Heat treatment of Panax ginseng C. A. Meyer at the temperature higher than that applied to the conventional preparation of red ginseng yielded a mixture of saponins with potent antioxidative properties. Thus, the methanol extract of heat-processed ginseng (designated as'NGMe') attenuated lipid peroxidation in rat brain homogenates induced by ferric ion or ferric ion plus ascorbic acid. Furthermore, the extract protected against strand scission in f Xl 74 supercoiled DNA Induced by UV photolysis of H2O2 and was also capable of scavenging superoxide generated in vitro by xanthine/xanthine oxidate or in differentiated human promyelocytic leukemia (HL-60) cells by the tumor promoter,12-0-tetvade- canoylphorbol-13-acetate (TPA). Since tumor promotion is closely linked to oxidative stress, we have determined possible anti-tumor promotional effects of NGMe on two-stage mouse skin tumorigenesis. Topical application of NGMe onto shaven backs of female ICR mice 10 min prior to TPA significantly ameliorated skin papillomagenesi s initiated by 7,12-dimethylbenz (a) anthracene (DMBA).'Likewise, TPA-induced epidermal ornithine decarboxylase activity and elevation of tumor necrosis factor-a were suppressed signifies%fly by NGMe pretreatment. NGMe topically applied onto surface of hamster buccal pouch 10 min before each topical application of DMBA inhibited oral carcinogenesis by 76olo in terms of multiplicity. Taken together, these results suggest that processed Panax ginseng C. A. Meyer has potential cancer chemopreventive activities.

• Journal of Pharmaceutical Investigation
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• 제36권1호
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• pp.1-9
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• 2006

Human solid tumors exhibit a multicellular resistance (MCR) resulting from limited drug penetration and decreased sensitivity of tumor cells when interacting with their microenvironments. Multicellular cultures represent solid tumor condition in vivo and provide clinically relevant data. There is little data on antitumor effect of paclitaxel (PTX) in multicellular cultures although its MCR has been demonstrated. In the present study, we evaluated antiproliferative effects of PTX in multicellular layers (MCL) of DLD-1 human colorectal carcinoma cells. BrdU labeling index (LI), thickness of MCL, cell cycle distribution and cellular uptake of calcein were measured before and after exposure to PTX at 0.1 to 50 ${\mu}M$ for 24, 48 and 72 hrs. BrdU LI and thickness of MCL showed a concentration- and time-dependent decrease and the changes in both parameters were similar, i.e., 34.2% and 40.6% decrease in BrdU LI and thickness, respectively, when exposed to $50\;{\mu}M$ for 72 hr. The DLD-1 cells grown in MCL showed increase in $%G_{0}/G_{1}$ and resistance to cell cycle arrest and apoptosis compared to monolayers. Calcein uptake in MCL did not change upon PTX exposure, indicating technical problems in multicellular system. Overall, these data indicate that antitumor activity of PTX may be limited in human solid tumors (a multicellular system) and MCL may be an appropriate model to study further pharmacodynamics of PTX.

• 대한본초학회지
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• 제21권2호
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• pp.37-46
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• 2006

Objectives : The purpose of this study was to investigate the effect of Dangguibohyultang (DB) and its combination (DB-I; Astragali membraneus BUNGE : Angelica gigas NAKAI=5:1, DB-II; Astragali membraneus BUNGE:Angelica gigas NAKAI=1:1, DB-III; Astragali membraneus BUNGE:Angelica gigas NAKAI=1:5,) on apoptosis in human colorectal adenocarcinoma HCT116 cells. Methods : To study the cytotoxic effect of methanol extract of DB-I, DB-II and DB-III on HCT116 cells, the cell viability was determined by XTT reduction method and ttypan blue exclusion assay. To confirm the induction of apoptosis, the cleavage of poly ADP-ribose polymerase (PARP), a substrate for caspase-3 and a typical sign of apoptosis, and the activation of procaspase-3, -8 and -9 were examined by western blot analysis. Furthermore, DB-induced apoptosis was confirmed by DNA fragmentation. The release of cytochrome C from mitochondria to cytosol, the level of Bcl-2 and Bax, and the expressions of Raf/MEK/ERK were examined by western blot analysis. Results : DB-I and DB-II reduced proliferation of HCT116 cells in a dose-dependent manner. DB-I and DB-II decreased procaspase-3, -8, -9 levels in a dose-dependent manner and induced the clevage of PARP. DB-I and DB-II also triggered the mitochondrial apoptotic signaling by increasing the release of cytochrome C from mitochondria to cytosol, decreasing of anti-apoptotic Bcl-2, and increasing of pro-apoptotic Bax. DB-I and DB-II decreased the activation of Ras/Raf/MEK/ERK cascade in a dose-dependent manner. Conclusion : These results suggest that DB-I and DB-II induce apoptosis via mitochondrial pathway in HCT116 cells. Furthermore, Raf/MEK/ERK cascade is involved in DB-induced apoptosis. These results suggest that DB is potentially useful as a chemotherapeutic agent in human liver cancer.

• 생명과학회지
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• 제24권9호
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• pp.967-972
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• 2014

본 연구는 한국 고유의 전통주인 막걸리 제조시 생성되는 주박 추출물과 유기용매 분획물을 총 85종 분리하여, 이들의 대장암 세포주에 대한 항성장 활성과 활성기전을 연구하였다. 이들 중 세포생존율 연구결과에 따라 3가지 종류의 분획물(KSD-E1-3, KSD-E2-3, KSD-E4-3)을 선별하였다. 이 중 가장 활성이 높은 KSD-E1-3 분획물에 의한 유전체 수준에서의 유전자 발현변화를 확인하기 위하여 oligo DNA microarray 실험을 수행하였다. 그 결과, 2배이상 발현이 증가된 유전자들 가운데 항암 유전자인 NAG-1, ATF3, DDIT3 그리고 p21을 선별하여 추가 실험을 진행하였다. 세포생존율 결과에 따라 선별된 3가지 종류의 분획물(KSD-E1-3, KSD-E2-3 그리고 KSD-E4-3)에 의한 4가지 종류의 유전자의 발현을 확인한 결과, KSD-E1-3에 의하여 모든 유전자의 발현이 높게 증가되었다. KSD-E1-3에 의한 유전자의 발현이 전사조절인자 p53에 의존적인지 확인하고자 p53 null HCT116 세포주를 이용하여 RT-PCR한 결과, NAG-1, ATF3, 그리고 DDIT3 유전자 p53에 비의존적이었으며, 반면 p21 유전자는 p53에 의해서만 발현이 증가되는 것을 확인하였다. 이러한 연구 결과는 주박 추출물이 다양한 기작에 의해 항암 유전자의 발현을 유도함으로써 암 세포에 대한 항성장 활성을 보여주고 있음을 나타낸다.

• IMMUNE NETWORK
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• 제4권1호
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• pp.23-30
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• 2004

Background: A human orthologue of mouse S100A6-binding protein (CacyBP), Siah-1-interacting protein (SIP) had been shown to be a component of novel ubiquitinylation pathway regulating $\beta$-catenin degradation. The role of the protein seems to be important in cell proliferation and cancer evolution but the expression pattern of SIP in actively dividing cancer tissues has not been known. For the elucidation of the role of SIP protein in carcinogenesis, it is essential to produce monoclonal antibodies specific to the protein. Methods: cDNA sequence coding for ORF region of human SIP gene was amplified and cloned into an expression vector to produce His-tag fusion protein. Recombinant SIP protein and monoclonal antibody to the protein were produced. The N-terminal specificity of anti-SIP monoclonal antibody was conformed by immunoblot analysis and enzyme linked immunosorbent assay (ELISA). To study the relation between SIP and colon carcinogenesis, the presence of SIP protein in colon carcinoma tissues was visualized by immunostaining using the monoclonal antibody produced in this study. Results: His-tag-SIP (NSIP) recombinant protein was produced and purified. A monoclonal antibody (Korea patent pending; #2003-45296) to the protein was produced and employed to analyze the expression pattern of SIP in colon carcinoma tissues. Conclusion: The data suggested that anti-SIP monoclonal antibody produced here was valuable for the diagnosis of colon carcinoma and elucidation of the mechanism of colon carcinogenesis.

• 한국균학회지
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• 제29권1호
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• pp.61-66
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• 2001

곤충기생성 균류인 번데기동충하초는 각종 약리활성을 나타내는 것으로 유명하다. 국내에서 인공재배한 번데기동충하초 자실체를 n-hexane으로 추출하여 항암성분을 실리카젤 칼럼크로마토그래피로 순수하게 분리하여 $^1H-NMR$과 $^{13}C-NMR$로 그 구조를 밝혀본 결과 ergosterol peroxide $(5{\alpha},\;8{\alpha}-epidioxy-24(R)-methylcholesta-6,22-dien-3{\beta}-ol)$로 밝혀졌다. 분리한 crgosterol peroxide를 한국인의 암환자에서 분리한 각종 암세포에 대하여 항암작용을 측정한 결과, 3일 후에 위암세포주인 SNU-1 암세포에 대하여 가장 강한 항암작용을 나타내었다. 한국인의 암환자에서 유래한 위암세포인 SNU-1, 간암세포인 SNU-354 및 직장암세포인 SNU-C4 암세포 등에 대한 6일 후에 ergosterol peroxide의 50% 성장억제농도는 각각 75.8, 39.7, $32.7{\mu}g/ml$이었다. 따라서 ergosterol peroxide 성분은 번데기동충하초의 항암성분중의 하나로 밝혀졌다.

• 대한방사성의약품학회지
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• 제1권2호
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• pp.118-122
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• 2015

Suicidal gene therapy is based on the transduction of tumor cells with "suicide" genes encoding for prodrug-activating enzymes that render target cells susceptible to prodrug treatment. Suicidal gene therapy results in the death of tumor with the expression of gene encoding enzyme that converts non-toxic prodrug into cytotoxic product. Cytochrome P450 4B1 (CYP4B1) activates 4-ipomeanol (4-IPO) or 2-aminoanthracene (2-AA) to cytotoxic furane epoxide and unsaturated dialdehyde intermediate.In this study, therapeutic effects of suicidal gene therapy with rabbit CYP4B1/2-AA or 4-IPO system were evaluated in HT-29 (human colon cancer cell). pcDNA-CYP4B1 vector was transfected into HT-29 by lipofection and stable transfectant was selected by treatment of hygromycin ($500{\mu}g/mL$) for 3 weeks. Reverse transcription polymerase chain reaction (RT-PCR) analysis was performed for confirmation of CYP4B1 expression in CYP4B1 gene transduced cell. The cytotoxic effects of CYP4B1 transduced cell were determined using dye-exclusion assay after treatment of 2-AA or 4-IPO for 96 hrs. Dye-exclusion assay showed that $IC_{50}$ of HT-29 and CYP4B1 transduced HT-29 was 0.01 mM and 0.003 mM after 4-IPO or 2-AA treatment at 96 hrs exposure, respectively. In conclusion, CYP4B1 based prodrug gene therapy probably have the potential for treatment of colorectal adenocarcinoma.