• 생명과학회지
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• 제17권2호통권82호
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• pp.179-184
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• 2007

단백질 티로신 kinase인 PTK6는 대부분의 유방암에서 과발현되며, 암세포의 증식만을 촉진하는데 역할을 한다. 이 연구에서 PTK6의 활성도메인을 초파리의 peptidoglycan recognition protein (PGRP) -LB 단백질을 퓨전파트너로 사용하여 바큘로바이러스 시스템을에서 과발현하는데 성공하였다. 우리는 PGRP-LB가 바큘로바이러스 시스템에서 잠재적으로 퓨전 단백질로 사용될 수 있는 가능성을 처음으로 발견하였다. 정제된 PTK6단백질은 기존의 박테리아에서 발현된 단백질보다 1.5배 높은 활성을 지녔다. 이 단백질은 PTK6의 분자기전 및 그것의 저해제 개발에 필수적인 결정 구조를 규명하는데 사용될 것이다.

• Biotechnology and Bioprocess Engineering:BBE
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• 제7권1호
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• pp.1-5
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• 2002

A fusion protein, consisting of a human epidermal growth factor (hEGF) as the recognition domain and human angiogenin as the toxin domain, can be used as a targeted therapeutic against breast cancer cells among others. The fusion protein was expressed as inclusion body in recombinant E. coli, and when the conventional, solution-phase refolding process was used the refolding yield was very low due to severe aggregation. It was probably because of the opposite electric charge at a neutral pH resulting from the vastly different pI values of each domain. The solid-phase refolding process that exploited the ionic interactions between ionic exchanger surface and the fusion protein was tried, but the adsorption yield was also very low, below $ 30\%$, regardless of the resins and pH conditions used. Therefore, to provide a higher ionic affinity toward the solid matrix, six lysine residues were tagged to the N-terminus of the hEGF domain. When heparin-Sepharose was used as the matrix, the adsorption capacity increased 2.5-3 times to about $88\%$. Besides the intrinsic affinity of angiogenin to heparin, the poly-lysine tag provided additional ionic affinity. And the subsequent refolding yield increased nearly 13-fold, from ca. $4.8\%$ in the conventional refolding of the untagged fusion protein to $63.6\%$. The process was highly reproducible. The refolded protein in the column eluate retained RNase bioactivity of angiogenin.

• 한국식품저장유통학회지
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• 제11권4호
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• pp.461-467
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• 2004

장기간 자연숙성시킨 전통된장의 항산화 효과는 숙성기간이 길어질수록 대체로 감소되었으며, 숙성기간 1년 된장에서 methanol 분획물이 가장 우수하였으며, 다음으로 hexane과 물 분획물의 순이었다. 지용성 및 수용성 추출물은 숙성기간이 길어질수록 항산화 효과가 점진적으로 오히려 증가되었으나, 분획물에 비하여 낮은 수치를 나타내었다. 전통된장의 수소공여능은 수용성 색소 추출물과 methanol 및 butanol 분획물에서 대체적으로 높은 수치였으나, chloroform과 ethylacetate 분획물은 매우 낮았다. 전통된장의 항암효과에서 인체 폐암 세포주(A549)는 물 분획물에서 가장 높았고, 인체 유방암 세포주(MCF-7)는 methanol 분획물에서 가장 높은 수치를 나타내었으며, 특히 전통된장의 숙성기간이 길어질수록 암세포 성장억제효과는 높게 나타났다.

• Journal of Ginseng Research
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• 제36권2호
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• pp.169-175
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• 2012

Ginseng has been used as a traditional medicine for treatment of many diseases and for general health maintenance in people of all ages. Ginseng is also used to ameliorate menopausal systems. We investigated the estrogenic activity of Korean red ginseng (KRG) in a transient transfection system, using estrogen receptor (ER) and estrogen-responsive luciferase plasmids in MCF-7 cells. The extract activated both ER${\alpha}$ and ER${\beta}$. KRG modulated the mRNA levels of estrogen-responsive genes such as pS2 and ESR1 and decreased the protein level of ER${\alpha}$. In order to examine in vivo estrogenic activity of KRG, sixteen female Sprague-Dawley rats separated into four groups were studied for nine weeks: non-ovariectomized (OVX) rats treated with olive oil, OVX rats treated with olive oil, OVX rats treated with 17-${\beta}$-estradiol (E2) in olive oil, and OVX rats treated with KRG extract in olive oil. The experiments were repeated for three times and the data of twelve rats were combined. Body weight of OVX rats was greater than that of sham-operated control rats and was decreased by E2 treatment. Uterine weight increased after E2 treatment compared to OVX rats. However, no difference in body or uterine weight was observed with KRG intake. KRG induced reductions in total cholesterol, low density lipoprotein cholesterol/total cholesterol, high density lipoprotein cholesterol/total cholesterol, and low density lipoprotein cholesterol/high density lipoprotein cholesterol, but not to the same degree as did E2 intake. These results show that KRG does contain estrogenic activity as manifested by in vitro study but the activity is not strong enough to elicit physiological responses.

• Archives of Pharmacal Research
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• 제23권4호
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• pp.338-343
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• 2000

The effects of methanol extract of Rubus crategifolius roots and its solvent fractions were investigated on the proliferation of MCF-7 human breast carcinoma cells. The methanol extract inhibited the proliferation of MCF-7 cells in a concentration dependent manner. Moreover, their methanol soluble (W-M) fraction had the greatest inhibitory effect on the growth of MCF-7 cells. To evaluate whether the W-M fraction affects on the cell cycle of MCF-7 cells, cells treated with this fraction were analyzed with flow cytometry. The W-M fraction increased $G_0$/$G_1$phase after 24 h-treatment and induced apoptosis after 48 h-treatment. The hallmark of apoptosis, DNA fragmentation, also appeared by W-M fraction after 48 h-treatment. Furthermore, the methanol extract and its W-M fraction inhibited the activity of the topoisomerase 1 enzyme in the relaxation assay, From these results, their W-M fraction as well as methanol extract of R. crategifolius roots are necessary for further studies as a potent inhibitor of the growth of cancer cells.

• 한국환경성돌연변이발암원학회지
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• 제24권4호
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• pp.180-188
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• 2004

CYP1A1 is known to be inducible by xenobiotic compouds such as polyciclic aromatic hydrocarbons(PAHs) and 2,3,7,8-tetrachloro-dibenzo-p-dioxin(TCDD). These chemicals have been identified worldwide and can have a significant impact on the human health and well being of human and wildlife. Given these issues, the detection and quantification of these chemicals in biological, environmental and food samples is important. We investigated the effect of dietaty flavonoid, such as CYP1A1 promoter activity, 7-ethoxyresorufin-O-deethylase(EROD) activity and CYP1A1 mRNA expression induced by benzo(k)fluoranthene(B(k)F) in MCF-7 cells. Based on the three criteria of frequency of occurrence in the environment, toxicity and potential exposure to humans, B(k)F is one of the top-listed PAHs. We found that B(k)F significantly up-regulates the level of CYP1A1 promoter activity, EROD and CYP1A1 mRNA. when cells were treated with daidzein inhibited the B(k)-induced CYP1A1 prompter activity and mRNA level at high concentration. But daidzein exhibited stimulatory effects B(k)F-induced CYP1A1 promoter activity and mRNA level at low concentration. Overall, results from these studies demonstrate flavonoids might interfere the action of B(k) with AhR system to stimulate CYP1A1 gene expression.

• 한국환경성돌연변이발암원학회지
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• 제24권4호
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• pp.189-197
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• 2004

We investigated the effect of dietaty flavonoid, such as CYP1A1 promoter activity, 7-ethoxyresorufin-O-deethylase(EROD) activity and CYP1A1 mRNA expression induced by benzo(k)fluoranthene(B(k)F) in MCF-7 cells. Based on the three criteria of frequency of occurrence in the environment, toxicity and potential exposure to humans, B(k)F is one of the top-listed PAHs. We found that B(k)F significantly up-regulates the level of CYP1A1 promoter activity, EROD and CYP1A1 mRNA. When cells were treated with morin alonem, it was not changed that EROD and CYP1A1 mRNA, compared to that of control. However, morin inhibited the B(k)-induced CYP1A1 prompter activity and mRNA level at high concentration. But morin exhibited stimulatory effects B(k)F-induced CYP1A1 promoter activity and mRNA level at low concentration. Overall, results from these studies demonstrate morin might interfere the action of B(k) with AhR system to stimulate CYP1A1 gene expression. CYP1A1 is known to be inducible by xenobiotic compouds such as polyciclic aromatic hydrocarbons(PAHs) and 2,3,7,8-tetrachloro-dibenzo-p-dioxin(TCDD). These chemicals have been identified worldwide and can have a significant impact on the human health and well being of human and wildlife. Given these issues, the detection and quantification of these chemicals in biological, environmental and food samples is important.

• BMB Reports
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• 제38권3호
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• pp.300-306
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• 2005

Tamoxifen citrate is an anti-estrogenic drug used for the treatment of breast cancer. It showed a degree of hepatic carcinogenesis, when it used for long term as it can decrease the hexose monophosphate shunt and thereby increasing the incidence of oxidative stress in liver rat cells leading to liver injury. In this study, a model of liver injury in female rats was done by intraperitoneal injection of tamoxifen in a dose of 45 mg/kg body weight for 7 successive days. This model produced a state of oxidative stress accompanied with liver injury as noticed by significant declines in the antioxidant enzymes (glutathione-S-transferase, glutathione peroxidase and catalase) and reduced glutathione concomitant with significant elevations in TBARS (thiobarbituric acid reactive substance) and liver transaminases; sGPT (serum glutamate pyruvate transaminase) and sGOT (serum glutamate oxaloacetate transaminase) levels. The oral administration of dimethyl dimethoxy biphenyl dicarboxylate (DDB) in a dose of 200 mg/kg body weight daily for 10 successive days, resulted in alleviation of the oxidative stress status of tamoxifen-intoxicated liver injury in rats as observed by significant increments in the antioxidant enzymes (glutathione-S-transferase, glutathione peroxidase and catalase) and reduced glutathione concomitant with significant decrements in TBARS and liver transaminases; sGPT and sGOT levels. The administration of DDB before tamoxifen intoxication (as protection) is more little effective than its curative effect against tamoxifen-induced liver injury. The data obtained from this study speculated that DDB can mediate its biochemical effects through the enhancement of the antioxidant enzyme activities and reduced glutathione level as well as decreasing lipid peroxides.

• 생명과학회지
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• 제13권6호
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• pp.799-804
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• 2003

천연물에 포함되어 있는 에스트로겐 효과를 가지는 성분의 생체 내에서의 직접적인 효과를 검정하기 위하여, 에스트로겐 수용체를 발현하는 것으로 알려진 인체 유방암 세포주 MCF7에 에스트로겐에 반응성을 나타내도록 고안된 CAT 리포터 플라스미드를 도입한 in vitro 검출시스템을 사용하여 에스트로겐 활성을 측정하였다. 이미 여러 분야에서 그 기능성의 연구가 활발히 진행되고 있는 광합성 조류인 스피루리나(spirulina)와 파래 등의 해양식물을 대상으로 에스트로겐 반응 리포터 시스템을 이용하여 에스트로겐 활성을 측정하였다. 그 결과, 스피루리나 에탄올 추출물의 CAT 활성은 $500\mug/ml\; 와\; 50 \mug/ml$의 농도에서 표준물질인 $17\beta-estradiol$의 농도 $10^{-8}$ M과 비슷한 정도의 에스트로겐 활성 효과를 나타내었고, $5\mug/ml$의 농도에서는 표준물질인 $17\beta-estradiol$의 농도 $10^{-10}$ M과 비슷한 정도의 에스트로겐 활성 효과를 나타내었다. 하지만 파래 추출물의 경우에는 에탄올을 처리한 대조군과 비교하여 유의한 CAT활성 변화를 나타내지 않았다 한편, 불가사리와 새우 등의 해양동물을 대상으로 한 실험에서는 에탄올을 처리한 대조군과 비교하여 유의한 CAT 활성 변화를 나타내지 않았다. 이 연구 결과로 광합성 조류인 스피루리나에 에스트로겐 활성을 효과적으로 나타내는 생리활성성분이 포함되어 있을 수 있다는 가능성을 확인하였다.

• 대한방사성의약품학회지
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• 제1권2호
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• pp.130-136
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• 2015

We evaluate the influence of MR contrast agent on positron emission tomography (PET) image using phantom, animal and human studies. Phantom consisted of 15 solutions with the mixture of various concentrations of Gd-based MR contrast agent and fixed activity of [$^{18}F$]FDG. Animal study was performed using rabbit and two kinds of MR contrast agents. After injecting contrast agent, CT or MRI scanning was performed at 1, 2, 5, 10, and 20 minutes. PET image was obtained using clinical PET/CT scan, and attenuation correction was performed using the all CT images. The values of HU, PET activity and MRI intensity were obtained from ROIs in each phantom and organ regions. In clinical study, patients (n=20) with breast cancer underwent sequential acquisitions of early [$^{18}F$]FDG PET/CT, MRI and delayed PET/CT. In phantom study, as the concentration increased, the CT attenuation and PET activity also increased. However, there was no relationship between the PET activity and the concentration in the clinical dose range of contrast agent. In animal study, change of PET activity was not significant at all time point of CT scan both MR contrast agents. There was no significant change of HU between early and delayed CT, except for kidney. Early and delayed SUV in tumor and liver showed significant increase and decrease, respectively (P<0.05). Under the condition of most clinical study (< 0.2 mM), MR contrast agent did not influence on PET image quantitation.