• Asian Pacific Journal of Cancer Prevention
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• 제15권2호
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• pp.929-935
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• 2014

Two types of nanosized titanium dioxide, anatase ($anTiO_2$) and rutile ($rnTiO_2$), are widely used in industry, commercial products and biosystems. $TiO_2$ has been evaluated as a Group 2B carcinogen. Previous reports indicated that $anTiO_2$ is less toxic than $rnTiO_2$, however, under ultraviolet irradiation $anTiO_2$ is more toxic than $rnTiO_2$ in vitro because of differences in their crystal structures. In the present study, we compared the in vivo and in vitro toxic effects induced by $anTiO_2$ and $rnTiO_2$. Female SD rats were treated with $500{\mu}g/ml$ of $anTiO_2$ or $rnTiO_2$ suspensions by intra-pulmonary spraying 8 times over a two week period. In the lung, treatment with $anTiO_2$ or $rnTiO_2$ increased alveolar macrophage numbers and levels of 8-hydroxydeoxyguanosine (8-OHdG); these increases tended to be lower in the $anTiO_2$ treated group compared to the $rnTiO_2$ treated group. Expression of $MIP1{\alpha}$ mRNA and protein in lung tissues treated with $anTiO_2$ and $rnTiO_2$ was also significantly up-regulated, with $MIP1{\alpha}$ mRNA and protein expression significantly lower in the $anTiO_2$ group than in the $rnTiO_2$ group. In cell culture of primary alveolar macrophages (PAM) treated with $anTiO_2$ and $rnTiO_2$, expression of $MIP1{\alpha}$ mRNA in the PAM and protein in the culture media was significantly higher than in control cultures. Similarly to the in vivo results, $MIP1{\alpha}$ mRNA and protein expression was significantly lower in the $anTiO_2$ treated cultures compared to the $rnTiO_2$ treated cultures. Furthermore, conditioned cell culture media from PAM cultures treated with $anTiO_2$ had less effect on A549 cell proliferation compared to conditioned media from cultures treated with $rnTiO_2$. However, no significant difference was found in the toxicological effects on cell viability of ultra violet irradiated $anTiO_2$ and $rnTiO_2$. In conclusion, our results indicate that $anTiO_2$ is less potent in induction of alveolar macrophage infiltration, 8-OHdG and $MIP1{\alpha}$ expression in the lung, and growth stimulation of A549 cells in vitro than $rnTiO_2$.

• 한국식품위생안전성학회지
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• 제25권3호
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• pp.269-277
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• 2010

본 연구에서는 azoxymethane (AOM)과 dextran sodium sulfate (DSS)로 유도된 대장 발암과정에 대한 셀레늄의 방어 효과를 조사하였다. 셀레늄 결핍(0.02 ppm Se), 정상(0.1 ppm Se), 과다(0.5 ppm Se)사료를 12주간 식이로 급여하여 혈액검사와 대장암 발생의 초기단계인 aberrant crypt foci (ACF)수를 측정했으며, 암 발생율을 조사하였다. ICP-AES를 사용하여 간의 셀레늄 농도를 측정하였으며, 또한 셀레늄포함 항산화효소인 glutathione peroxidase (GPx) 활성을 알아보았다. 또한 TUNEL assay와 PCNA, $\beta$-catenin에 대한 면역조직 염색을 수행하였다. ACF 수 및 종양 발생률에 있어서, 셀레늄과다사료를 급여한 군이 정상셀레늄사료를 급여한 군보다 낮았으며, 셀레늄결핍사료를 급여한 군은 오히려 ACF 수 및 종양 발생률이 높았다. GPx 활성은 셀레늄의 섭취가 과다한 군에서 높게 나타났으며, 이 때, TUNEL에서 apoptotic positive cell이 증가하는 것을 확인했다. 또한 셀레늄의 섭취가 과다한 군에서 PCNA와 $\beta$-catenin의 발현이 감소됨을 볼 수 있었다. 본 마우스 모델실험에서 셀레늄은 여러 기전에 의해 대장암 발생을 억제할 수 있을 것으로 사료된다.

• 한국식품영양과학회지
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• 제39권1호
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• pp.54-63
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• 2010

Flavonoid 계열의 한 종류인 baicalin은 항염증, 항암, 항바이러스, 항세균 등의 효능을 가진다. 본 연구진은 선행연구를 통한 이전의 보고에서 baiclain이 adipogenesis pathway(지방세포 형성 경로)의 anti-adipogenic(지방세포 형성억제)과 pro-adipogenic(지방세포 형성 유도) factor들을 조절함으로써 비만 및 adipogenesis를 억제함을 밝혔다. 본 연구에서는, microarray 기술을 이용하여 3T3-L1 세포에서 baiclain이 유도하는 지방세포 형성 억제 효과에 대한 분자적 기작을 보다 상세하게 연구하고자 하였다. 지방세포의 분화 시간(0일, 2일, 4일 및 7일)과 분화 시 baicalin의 처리 유무에 따라 유전자 발현 양상을 분석하기 위해 해당 시료들을 microarray에 적용하였다. Microarray 결과로부터 2배이상의 변화가 있는 3972개의 유전자를 확보하였다. 그 유전자들의 발현 양상을 좀 더 자세히 살펴보기 위해 hierarchical clustering 분석을 진행하였고 그 결과로 20개의 cluster를 분류할 수 있었다. 그들 중 4개의 cluster는 분화의 전반적인 기간에서 baicalin의 첨가에 의해 뚜렷하게 상승(cluster 8과 cluster 10)하거나 반대로 뚜렷하게 감소(cluster 12와 cluster 14)하는 양상을 보였다. Cluster 8과 cluster 10에는 CHOP(CCAAT/enhancer-binding protein homologous protein), INSIG1(insulin induced gene 1), WISP2(WNT1 inducible signaling pathway protein 2), ADM(adrenomedullin), CCND2(cyclin D2), GRN(granulin) 및 TGFB3(transforming growth factor, beta 3)과 같은 세포 증식과 지방세포 형성 억제를 상승시키는 유전자들이 다수 포함되었다. 반대로 cluster 12와 cluster 14에는 세포 증식 억제, 세포 주기 억제 및 세포 성장 억제와 연관되거나 지방세포를 유도하는 유전자인 LTA(lympotoxin A), ACADSB(acyl-Coenzyme A dehydrogenase, short/branched chain), HMGCS2(3-hydroxy-3-methylglutaryl-Coenzyme A synthase 2), IGFBP7(insulin-like growth factor binding protein 7), MERTK(c-merproto-oncogene tyrosine kinase), RASSF2(ras association(RalGDS/AF-6) domain family 2), RHOU(ras homolog gene family, member U) 및 SESN1(sestrin1) 등이 포함되었다. 결론적으로 baicalin은 세포 증식 및 지방세포 형성과 연관된 유전자들을 조절함으로써 지방세포의 분화를 억제하는 것으로 사료된다. 이러한 결과는 baicalin이 유도하는 지방세포 형성 억제 및 비만 억제 효과의 분자적 기작에 대한 중요한 정보를 제시한다.

• 한국미생물·생명공학회지
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• 제42권1호
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• pp.58-66
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• 2014

본 연구에서는 브로콜리의 비가식 부위의 효율적인 이용 및 브로콜리의 가공식품 개발 연구의 일환으로 브로콜리의 주요 가식부위인 꽃송이와 비가식 부위인 줄기 부분을 대상으로 각각 에탄올 추출물 및 이들의 순차적 유기용매 분획물을 조제하여 이들의 유용성분 분석 및 항산화, 항균, 대장암 세포 생육억제능을 평가하였다. 먼저 성분 분석결과 브로콜리 줄기는 꽃송이보다 수용성 당류를 많이 포함하고 있으며, 꽃송이는 상대적으로 지용성 물질을 많이 포함하고 있었고, 총 플라보노이드 및 총 폴리페놀은 꽃송이가 줄기보다 1.50-1.99배 높은 함량을 나타내었다. 분획물의 총 플라보노이드 함량은 줄기 및 꽃송이의 ethylacetate 분획이 9.45 mg/g 및 42.01 mg/g으로 가장 높게 나타났으며, 총 폴리페놀 함량도 꽃송이 ethylacetate 분획이 181.12 mg/g으로 가장 높게 나타났다. 항산화 활성 평가결과, 브로콜리 꽃송이 에탄올 추출물의 ethylacetate 분획에서 강력한 DPPH 음이온 소거능, ABTS 양이온 소거능 및 환원력을 확인하였으며, nitrite 소거능의 경우에는 줄기의 n-hexane 분획이 강력한 활성을 나타내었다. 항균 활성 평가에서는, 꽃송이의 ethylacetate 분획에서 실험에 사용된 그람 양성 및 그람 음성 세균 모두에서 강력한 항균력을 나타내었다. 대장암 세포성장 억제활성 평가에서는, 줄기 및 꽃송이에서 모두 n-hexane 분획 및 ethylacetate 분획에서 우수한 생육억제 활성을 나타내었으며, 줄기의 경우 n-hexane 분획에서 세포 생존율 $18.4{\pm}1.2%$, 꽃송이에서는 n-butanol 분획에서 세포 생존율 $6.9{\pm}2.3%$를 나타내어 가장 강력한 활성이 나타났다. 본 연구결과는 브로콜리의 다양한 부위로부터 관련 활성물질의 탐색, 정제를 통한 식의약품 소재 개발 연구가 가능함을 제시하고 있다.

• 생명과학회지
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• 제23권9호
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• pp.1118-1125
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• 2013

감귤류에 많이 함유되어 있는 naringenin은 항암화학요법제로서 중요한 가능성을 가지고 있으나 항암활성에 대한 분자생물학적 기전에 대해서는 명확히 밝혀져 있지 않다. 본 연구에서는 인체 혈구암세포인 U937 세포에서 naringenin이 유발하는 항암효과 및 항암기전을 조사하였다. Naringenin 처리에 의한 U937 세포의 증식억제는 apoptosis 유발과 연관성이 있었으며, 이러한 현상은 caspases 활성화와 밀접한 관련이 있었다. 그러나 pan-caspase inhibitor인 z-VAD-fmk의 선처리에 의하여 U937 세포에서 naringenin이 유발하는 apoptosis가 억제되는 것으로 나타났으므로 caspases가 apoptosis 유발의 중요한 조절자라는 것을 알 수 있었다. 또한 U937 세포에 naringenin을 처리하였을 경우 pro-apoptotic Bcl-2 및 anti-apoptotic Bax의 발현에는 아무런 변화가 나타나지는 않았지만 Bcl-2가 과발현된 U937/Bcl-2 세포에서 naringenin에 의한 apoptosis가 억제되었다. 하지만 small-molecule Bcl-2 inhibitor인 HA14-1 및 naringenin을 같이 처리하였을 경우에는 XIAP 발현감소, Bid 단편화 및 caspase-3 활성화를 통하여 다시 apoptosis가 유발되었다. 따라서 HA14-1 및 naringenin에 의한 apoptosis 상승효과는 death receptor-mediated apoptosis pathway를 경유한다는 것을 제시하는 결과이다.

• 생명과학회지
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• 제19권2호
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• pp.249-255
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• 2009

Esculetin, a coumarin compound, has been known to inhibit proliferation and induce apoptosis in several types of human cancer cells. However, the molecular mechanisms involved in esculetin-induced apoptosis are still uncharacterized in human leukemia cells. In this study, we have investigated whether esculetin exerts anti-proliferative and apoptotic effects on human leukemia U937 cells. It was found that esculetin could inhibit cell viability in a time-dependent manner, which was associated with the induction of apoptotic cell death such as increased populations of apoptotic- sub G1 phase. Apoptosis of U937 cells by esculetin was associated with an inhibition of Bcl-2/Bax binding activity, formation of tBid, down-regulation of X-linked inhibitor of apoptotic protein (XIAP) expression, and up-regulation of death receptor 4 (DR4) and FasL expression. Esculetin treatment also induced the degradation of ${\beta}$-catenin and DNA fragmentation factor 45/inhibitor of caspase-activated DNase (DFF45/ICAD). Furthermore, a caspase-3 specific inhibitor, z-DEVD-fmk, significantly inhibited sub-G1 phase DNA content, morphological changes and degradation of ${\beta}$-catenin and DEE45/ICAD. These results indicated that a key regulator in esculetin-induced apoptosis was caspase-3 in human leukemia U937 cells.

• 한국식품저장유통학회:학술대회논문집
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• 한국식품저장유통학회 2003년도 춘계총회 및 제22차 학술발표회
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• pp.22-39
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• 2003

Biological activity of phenolic compounds in seeds and leaves of safflower (Carthamu tinctorius L.) were evaluated using several in vitro and in vivo assays. Six phenolic constituents were isolated from the seeds and identified as N-feruloylserotonia, N- (p-coumaroyl)serotonin, matairesinol, 8′-hydroxyarctigenin, acacetin 7-O-$\beta$-D-glucoside (tilianine) and acacetin. Six phenolic compounds exhibited considerable antioxidative activity, and especially two serotonins showed potent DPPH radical scavenging activity and antiperoxidative activity against rat liver microsomal lipid peroxidation induced by the hydroxyl radical generated via a Fenton-type reaction. Additionally, six phenolic compounds possessed comparable cytotoxicity against three cancer cells, Hela cell, MCF-7 and HepG2 cell, and particularly acacetin and its glycosides had the most potent cytotoxicity. Moreover, we found that feeding safflower seeds attenuated bone loss, and lowered levels of plasma and liver lipids in ovariectomized rats. Serotonins, lignans and flavones stimulated proliferation of the osteoblast-like cells in a dose-dependent manner (10$^{-15}$ ~10$^{-6}$ M), as potently as E$_2$ (17$\beta$-estradiol). Particularly, serotonins were mainly responsible for bone-protecting and lipid lowering effects in ovariectomized rats. Meanwhile, eight flavonoids, including a novel quercetin-7-O-(6"-O-acetyl)-$\beta$-D-glucopyranoside and seven kown flavonoids, luteolin quercetin, luteolin 7-O-$\beta$-D-glucopyranoside, luteolin-7-O-(6"-O-acetyl)-$\beta$-D-gluco-pyranoside, quercetin 7-O- -glucopyranoside, acacetin 7-O-$\beta$-D-glucuronide and apigenin-6-C-$\beta$-D-glucopyranosyl-8-C-$\beta$-D-glucopyranoside were first isolated and identified from safflower leaf. Among these flavonoids, luteolin-acetyl-glucoside and $\beta$quercetin- acetyl-glucoside showed potent antioxidative activities against 2-deoxyribose degradation and lipid peroxidation in rat liver microsomes. Luteolin, quercetin and their corresponding glycosides also exhibited strong antioxidative activity, while acacetin glucuronide and apigenin-6, 8-di-C-glucoside were relatively less active. Finally, changes in phenolic compositions were also determined by HPLC in the safflower seed and leaf during growth stages and roasting process to produce standardized supplement powerds. These results suggest that phenolic compounds in the roasted safflower seed and leaf may be useful as potential sources of therapeutic agents against several pathological disorders such as carcinogenesis, atherosclerosis and osteoporosis.

• 한국식품저장유통학회지
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• 제12권5호
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• pp.496-500
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• 2005

홍경천(Rhodiola dumulosa)을 기능성 식품소재로 개발하기 위하여 홍경천(뿌리)을 80% 메탄올로 열수 추출하여 용매 분획한 후 이들에 대한 항산화, 암세포 성장 억제 및 대식세포의 면역활성 등과 같은 기능성을 조사하였다. 홍경천 메탄올 추출물의 헥산, 에틸 아세테이트, 부탄올, 메탄올 및 물 분획물의 수소공여능은 100 ${\mu}g/mL$의 농도에서 각각 28.30, 53.21, 35.48, 42.64 및 21.14% 로서 에틸아세테이트 분획물이 가장 높았으며, 이는 합성 항산화제인 BHT와 비슷한 항산화 효과를 나타내었다. A549 및 SW480 와 같은 암세포에 용매 분획물 1, 10 및 100 ${\mu}g/mL$의 농도로 48시간 처리한 결과 에틸아세테이트 분획물이 다른 분획물에 비하여 암세포주의 성장을 강하게 억제하였으며, 그 활성은 농도 의존적으로 나타났다. 또한 100 ${\mu}g/mL$의 농도로 48시간 동안 에틸아세테이트 분획물을 암세포에 처리시 처리구에서의 암세포는 뚜렷한 세포수의 감소와 함께 심한 형태학적 변화가 관찰되었다. 대식세포주(RAW 264.7)에 1, 10 및 50 ${\mu}g/mL$의 농도로 용매 분획물을 처리한 후 NO의 생성량을 측정한 결과 10 μg/mL 이상의 농도에서 메탄올 분획물에서 NO 생성을 강하게 유도하였다. 따라서 본 결과는 홍경천의 기능성 식품소재로의 활용 가능성을 시사한다.

• 대한바이러스학회지
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• 제28권1호
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• pp.71-78
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• 1998

Vaccinia virus is the prototype orthopoxvirus that has been used as a vaccine strain for small pox. This virus has been used to express a variety of cellular and viral genes in mammalian cells at high levels. Interleukin-4 (IL-4) has been found to stimulate the proliferation of T cells and enhance the cytolytic activity of cytotoxic T lymphocytes. To test the immunotherapeutic potential of IL-4 delivered in vivo by poxvirus, a recombinant vaccinia virus expressing the murine IL-4 gene (RVVmIL-4) was constructed. A high level of IL-4 production was confirmed by infecting HeLa cells and measuring IL-4 in cell culture supernatant by ELISA. As a tumor model, two cell lines were used; the murine T leukemic line P388 and the murine breast cancer line TS/A. CDF1 mice were intraperitoneally inoculated with $1\;{\times}\;10^5$ cells of P388. Mice were injected at the same site with $5\;{\times}\;10^5\;PFU$ of recombinant vaccinia virus; first, 3 days after the injection of tumor cells and thereafter once every week for 3 weeks. Intraperitoneal injections of RVVmIL-4 significantly prolonged the survival time of mice inoculated with tumor cells. All mice injected with RVVmIL-4 remained alive for 30 days after the postinoculation of tumor cells, while 100% and 70% of the animals injected with saline or wild type vaccinia virus died, respectively. In another tumor model using TS/A, tumor was established by subcutaneously inoculating $2{\times}10^5$ tumor cells to BALB/c mice. After tumor formation was confirmed on day 4 in all mice, $5\;{\times}\;10^6\;PFU$ of RVVmIL-4 was inoculated subcutaneously three times, once every week for 3 weeks. The TS/A tumor was eradicated in two of the nine mice. Seven of the nine mice treated with RVVmIL-4 developed a tumor, but tumor growth was significantly delayed compared to those treated with saline or wild type vaccinia virus. These results indicate that recombinant vaccinia viruses may be used as a convenient tool for delivering immunomodulator genes to a variety of tumors.

• Molecular & Cellular Toxicology
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• 제5권2호
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• pp.126-132
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• 2009

The usage and types of chemicals are advancing, specializing, large-scaled increasing, and new chemical exposed workers are concerning to occupational disease. The generation of reactive oxygen in the body from carcinogen, mutation and DNA damage in cancer is protected by natural antioxidants (phytochemicals) with antimutagenic effect. There were many reports of ginsenoside Rb$_1$, Rg$_1$ grievances of the genetic mutation to suppress the effect confirm the genetic toxicity test with chromosomal aberration test and the Comet (SCGE) assay confirmed the suppression effect occurring chromosomal DNA damage. We had wanted to evaluate the compatibility and sensitivity between the chromosomal aberration (CA) test and the Comet assay. We used the CA test and Comet assay to evaluate the anti-genotoxicity of ginsenoside Rb$_1$ and Rg$_1$, in CHO-K1 (Chinese hamster ovary fibroblast) cell in vitro, composed negative control (solvent), positive control (benzo[a]pyrene), test group (carcinogen+variety concentration of ginsenoside) group. The positive control was benzo[a]pyrene (50 $\mu$M), well-known carcinogen, and the negative control was the 1 % DMSO solvent. The test group was a variety concentration of ginsenoside Rb$_1$, Rg$_1$ with 10$^{-8}$%, 10$^{-6}$%, 10$^{-4}$%, 10$^{-2}$%, 1%, 10%. In chromo-somal aberration test, we measured the number of cells with abnormally structured chromosome. In Comet assay, the Olive tail moment (OTM) and Tail length (TL) values were measured. The ratio of cell proliferation was increased 8.3% in 10$^{-8}$%, 10$^{-6}$%, 10$^{-4}$%, 10$^{-2}$%, 1%, 10% Rb$_1$ treated groups, and increased 10.4% in 10$^{-10}$%, 10$^{-8}$%, 10$^{-6}$%, 10$^{-4}$%, 10$^{-2}$%, 1% Rg$_1$ treated groups. In the CA test, the number of chromosomal aberration was decreased all the Rb$_1$ and Rg$_1$ treated groups. In the Comet assay, the OTM values were decreased in all the Rb$_1$ and Rg$_1$ treated groups. To evaluate the compatibility between CA and Comet assay, we compared the reducing ratio of chromosomal abnormalities with its OTM values, it was identified the antimutagenicity of ginsenoside, but it was more sensitive the CA test than the Comet assay. Ginsenoside Rb$_1$ and Rg$_1$ significantly decrease the number of cells with chromosomal aberration, and decrease the extent of DNA migration. Therefore, ginsenoside Rb$_1$, Rg$_1$ are thought as an antioxidant phytochemicals to protect mutagenicity. The in vitro Comet assay seems to be less sensitive than the in vitro chromosomal aberration test.