• 생명과학회지
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• 제25권8호
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• pp.849-860
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• 2015

본 연구에서는 한약재로 널리 사용되는 건칠, 유근피 및 신석 추출물의 항암 활성을 조사하였다. 생쥐 유래 정상세포(RAW 264.7 대식세포 및 C2C12 근아세포)에서는 건칠, 유근피 및 신석 단독 및 복합 처리에 의하여 유의적인 세포생존율의 억제 현상은 관찰 할 수 없었다. 그리고 건칠, 유근피 및 신석의 복합 처리는 단독 처리군에 비하여 AGS 위암세포의 생존력을 유의적으로 억제하였으나, 폐암(A549), 대장암(HCT116), 간암(Hep3B) 및 방광암(T24) 세포에서는 그 효과가 미비하였다. 아울러 이러한 AGS 위암세포 선택적 생존 억제력은 apoptosis 유도와 밀접한 연관성이 있음을 염색질의 응축 현상, DNA 단편화 및 annexin-V 염색에 의한 flow cytometry 분석을 통하여 확인하였다. 건칠, 유근피 및 신석의 복합 처리는 Fas 및 Fas legand의 발현을 증가시켰으며, XIAP, cIAP-1 및 survivin과 같은 IAP family 단백질과 anti-apoptotic Bcl-xL의 발현은 저하시켰다. 복합 처리는 또한 mitochondrial membrane potential의 손실과 caspases (-3, -8 및 -9)의 활성에 PARP 단백질의 분절화를 유도하였다. 그러나 이러한 복합 처리에 의한 AGS 세포에서 관찰된 세포독성 및 apoptosis 유도 효과는 pan-caspases inhibitor인 z-VAD-fmk의 선처리에 의하여 차단되었다. 이상의 결과는 건칠, 유근피 및 신석의 복합 처리에 의한 AGS 위암세포 선택적 apoptosis 유도가 caspase 의존적으로 일어나고 있음을 보여주는 결과이며, in vivo 모델을 이용한 후속 연구가 진행되어야 할 것이다.

• 대한골관절종양학회지
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• 제5권1호
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• pp.1-8
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• 1999

A single fraction of 50 Gy extracorporeal irradiation, as a modality of limb-sparing operation, has been used to achieve tumor necrosis in osteosarcoma. Although this modality of radiation therapy preserving the mobility of a joint is commonly practiced, the precise knowledge on the radiobiological response of osteosarcoma cell has remained to be elucidated. We therefore observed whether a single high dose irradiation caused apoptosis in osteosarcoma cells and whether the commitment to apoptosis was associated with cell kinetics. We also investigated radiation dose response along the time course for development of apoptosis following single high dose irradiation. The morphologic change in apoptosis was observed by fluorescence with Hoechst 33258 and the degree and the fraction of cells by flow cytometry. Irradiation of osteosarcoma cells with 10, 30 and 50 Gy resulted in chromatin condensation and apoptotic body formation. The degree of apoptosis in osteosarcoma cells was $29.5{\pm}3.56%$, $39.9{\pm}4.83%$ at 24 and 48 hours after 10 Gy irradiation ; $41.1{\pm}3.93%$, $66.9{\pm}5.21%$ at 24 and 48 hours after 30 Gy irradiation ; and $48.0{\pm}3.69%$, $75.6{\pm}4.65%$ at 24 and 48 hours after 50 Gy irradiation. The fraction of cells in cell-cycle kinetic was $39.2{\pm}4.3%$ in G2/M, $22.1{\pm}4.65%$ in G1 at 24 hours after 10 Gy irradiation ; $51.0{\pm}4.3%$ in G2/M, $20.4{\pm}4.7%$ in G1 at 48 hours after 10 Gy irradiation ; $40.3{\pm}3.9%$ in G2/M, $26.1{\pm}4.7%$ in G1 at 24 hours after 30 Gy irradiation ; $59.2{\pm}3.9%$ in G2/M, $5.9{\pm}5.1%$ in G1 at 48 hours after 30 Gy irradiation ; and $44.3{\pm}4.2%$ in G2/M, $21.1{\pm}3.5%$ in G1 at 24 hours after 50 Gy irradiation. The fraction of cells at 48 hours after 50 Gy irradiation could not be observed because of irradiation induced cell death of most of cells. All values for irradiated cells showed accumulation in G2/M phase and reduction in G1 phase, irrespective of irradiation dose. The results suggest that a single fraction of high dose irradiation with 50 Gy results in accumulation of cells at G2/M phase, leading to apoptosis.

• 한국세라믹학회지
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• 제41권10호
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• pp.777-782
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• 2004

액상응결 공정법과 일축가압성형법으로 제조된 기판위에 전해질과 양극층을 스크린 인쇄법으로 구성한 후 열처리함으로써 최종크기가 $5\times5cm^2$인 SOFC 단전지를 제조하였다. 본 연구에서는 이들 단전지와 인코넬 합금으로 제조된 접속자 그리고 가스켓형의 밀봉재를 이용하여 스택을 구성하였다. 본 연구에 사용된 스택은 연료가스와 산화가스가 교차되는 형태의 가스채널을 가지며 가스매니폴드가 내부에 구성되어 있는 형태로 설계되었다. 제작된 3단 스택의 성능을 평가해 본격과 15W 정도의 최고출력을 나타내었는데 이는 단전지 출력성능으로부터 예측된 최고출력치의 $50\%$ 정도에 해당되는 출력이었다. 본 연구에서는 이러한 스택성능에 영향을 주는 조정인자들과 스택디자인 인자들에 대한 분석을 수행하였다.

• 대한한방종양학회지
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• 제4권1호
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• pp.177-197
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• 1998

This experimental study was carried out to evaluate the effect of Yipahnsan on angiogenic inhibition mechanism. This study investigates the effects of Yipahnsan on angiogenic inhibition mechanism evaluate cell adhesive inhibition effect, DNA fragmantaion analysis, nuclear condensation assay, FACScan analysis, angiogenic lumen formation assay, immunocytochemistry analysis, RT-PCR for mRNA expression, western blot analysis, confocal analysis for $Ca^{2+}influx$. The results were summarized as follows : 1. The cell adhesive inhibition ability was strongly increased from $5{\mu}g/ml$ on ECV304 cell line and ECVPAR cell line. 2. YY water extract caused $G_0/G_1$ arrest peak to existed on the ECV304 cell line. 3. YY water extract caused inhibition of proliferation and inducement of apoptosis on the collagen coated plate in ECV304 cell line. 4. YY water extract inhibited the lumen formation on the matrigel coated plate in ECV304 cell line. 5. YY water extract inhibited the expressions of LFA-1 and ELAM-1 on ECV304 cell line and ECVPAR cell line. 6. YY water extract inhibited the expressions of MMP-9 and uPA on ECV304 cell line and ECVPAR cell line. 7. YY water extract inhibited the expression of integrin ${\alpha}_v{\beta}_3$ on ECV304 cell line and ECVPAR cell line. 8. YY water extract decreased the change of $Ca^{2+}$ in intracellular on ECV304 cell line and ECVPAR cell line. According to the results, Yipahnsan showed to be a key antagonist of integrin ${\alpha}_v{\beta}_3$, and to be induction of apoptosis by p53 through flow cytometry. This report also demonstrated that expressions of MMP-9 and uPA were blocked under the angiogenesis model. Thus, we suggested that Yipahnsan blocks angiogenesis by inducing apoptosis in ECV304 and ECVPAR cell lines, and another oriental herbal medicine that treats qi-stagnation and blood-stasis type also has angiogenic inhibition effects.

• 대한한방종양학회지
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• 제4권1호
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• pp.17-36
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• 1998

This experimental study was carried out to evaluate the effects of Hwallakhyoreungdan on angiogenic inhibition mechanism. In order to investigate the effects of Hwallakhyoreungdan on angiogenic inhibition mechanism, MTT assay, cell adhesive inhibition effect, DNA fragmantaion analysis, Nuclear condensation assay, FACScan analysis, Angiogenic lumen formation assay, Immunocytochemistry analysis, RT-PCR for mRNA expression, Western blot analysis and Confocal analysis for $Ca^{2+}$ change were performed. The results were summarized as follows: 1. The cell adhesive inhibition ability was strong from $5{\mu}g/ml$. 2. The $G_0/G_1$ arrest peak was existed on ECV304 cell-line. 3. The cells on Collagen plate were inhibition of proliferation and inducement of apoptosis by HR water extract. 4. Angiogenic lumen formation was inhibited by HR water extract. 5. LFA-1 and ELAM-1's expression were inhibited by HR water extract. They are commenly participation on inflammation and tumor regeneration. 6. The expression of MMP-9 and uPA were inhibited by HR water extract. 7. The expression of integrin ${\alpha}_v{\beta}_3$ was inhibited by HR water extract. 8. The expression of intracellular molecule were successively inhibited by HR water extract therefore the proliferation of ECV304 cell line was stopped and apoptosis was induced. 9. The change of $Ca^{2+}$ was decreased by HR water extract it cause confusion of signal transduction pathway therefore it was take part in apoptosis. According to the results, Hwallakhyoreungdan showed to be a key antaonist of integrin ${\alpha}_v{\beta}_3$, and to be induction of apoptosis by p53 through flow cytometry. This report also demonstrated that expressions of MMP-9 and uPA was blocked under the angiogenesis model. Thus, we suggests that Hwallakhyoreungdan blocks angiogenesis by inducing apoptosis of ECV304 and ECVPAR cell lines and another oriental herbal medicine that treats blood-stasis type also has angiogenic inhibition effects.

• 대한약침학회지
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• 제19권1호
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• pp.37-44
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• 2016

Objectives: This study examined the relative efficacies of a derivative of betulinic acid (dBA) and its poly (lactide-co-glycolide) (PLGA) nano-encapsulated form in A549 lung cancer cells in vivo and in co-mutagen [sodium arsenite (SA) + benzo[a]pyrene (BaP)]-induced lung cancer in mice in vivo. Methods: dBA was loaded with PLGA nanoparticles by using the standard solvent displacement method. The sizes and morphologies of nano-dBA (NdBA) were determined by using transmission electron microscopy (TEM), and their intracellular localization was verified by using confocal microscopy. The binding and interaction of NdBA with calf thymus deoxyribonucleic acid (CT-DNA) as a target were analyzed by using conventional circular dichroism (CD) and melting temperature (Tm) profile data. Apoptotic signalling cascades in vitro and in vivo were studied by using an enzyme-linked immunosorbent assay (ELISA); the ability of NdBA to cross the blood-brain barrier (BBB) was also examined. The stage of cell cycle arrest was confirmed by using a fluorescence-activated cell-sorting (FACS) data analysis. Results: The average size of the nanoparticles was ~ 110 nm. Confocal microscopy images confirmed the presence of NdBA in the cellular cytoplasm. The bio-physical properties of dBA and NdBA ascertained from the CD and the Tm profiles revealed that NdBA had greater interaction with the target DNA than dBA did. Both dBA and NdBA arrested cell proliferation at G0/G1, NdBA showing the greater effect. NdBA also induced a greater degree of cytotoxicity in A549 cells, but it had an insignificant cytotoxic effect in normal L6 cells. The results of flow cytometric, cytogenetial and histopathological studies in mice revealed that NdBA caused less nuclear condensation and DNA damage than dBA did. TEM images showed the presence of NdBA in brain samples of NdBA fed mice, indicating its ability to cross the BBB. Conclusion: Thus, compared to dBA, NdBA appears to have greater chemoprotective potential against lung cancer.

• 한국추진공학회지
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• 제19권1호
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• pp.61-67
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• 2015

연소기 시험 장치의 구축 시 고온의 연소 가스의 냉각은 중요한 설계요구조건이다. 물 분무(Water spray) 냉각 방식은 증발 과정에서 물의 잠열을 이용하므로, 효과적인 연소 가스 냉각이 가능하다. 본 연구에서는 연소기 시험 설비 구축 과정의 일환으로, 물 분무를 이용한 연소 가스의 냉각을 이해하기 위하여 연속방정식, 에너지 보존식과 포화 증기의 압력-온도 관계식을 이용한 1차원 해석을 수행하였다. 연소기 시험 장치에서 배출되는 고온, 고압의 연소 가스는 냉각수와의 혼합과정에서 배출가스의 온도가 낮아지며, 분무된 물의 일부는 기화하여 연소가스와 함께 배출되고, 일부는 다시 응축 되어 집수조로 모인다. 냉각수는 연소 가스의 온도를 낮춰주는 동시에, 증발된 증기는 연소기 내부의 압력을 증가시키므로 1차원 해석에서 증기의 압력-온도 관계식을 고려하여 해석을 수행하였다. 1차원 해석으로부터 연소가스의 적절한 냉각과 배기 덕트 내부의 압력의 지나친 상승을 피하기 위한 최적의 물 분무량을 확인하였으며, 물 분무 냉각 방식에 대한 물리적 이해를 얻을 수 있었다.

• 한국축산시설환경학회지
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• 제9권2호
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• pp.69-78
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• 2003

태양열을 동력원으로 하여 열에너지를 동력으로 변환, 물을 양수할 목적으로 저온 상변화물질인 펜탄을 작동물질로 하는 에너지변환장치를 제작하여 실험하였다. 장치는 각부의 크기를 그 기능과 상호작용 원리에 따라 논리에 맞게 최적 설계하였다. 장치의 제작 후 실험을 통하여 그 운전 특성을 분석하여 성능향상에 필요한 자료를 획득하고자 하였다. 작동물질인 펜탄을 가열하는 탱크 내부의 온도는 사이클 경과시간에 따라 약 $40­86^{\circ}C$ 범위에서 변동하고 있었으며, 물탱크내 온도 약 $23­24^{\circ}C$, 공기탱크 내 온도 $22­23.5^{\circ}C$ 범위에서 비교적 일정하게 유지되고 있었다. 응축기내의 온도와 냉각수출구 온도는 냉각수입구 온도수준에 따라 정의 상관관계로 변하고 있는 것을 알 수 있었으며, 또한 열 교환 능력도 냉각수 온도수준이 낮을수록 커진다는 것을 확인하였다. 물탱크 내 온도와 응축기내 온도가 상당히 차이가 나므로 물탱크와 응축기와의 연결거리를 최소화하고 연결파이프 크기를 큰 것으로 하여 내부물질이동 저항을 줄이는 것이 바람직하다는 것을 알 수 있었다. 실험 중 양수량은 1.6­2.4 liter로 나타났으며, 냉각시간의 수준에 따른 물탱크내의 흡입물높이 상승은 차이를 나타내지 않았다. 응축기로부터의 냉각수 배출파이프가 연장되지 않은 경우 냉각수 유량이 5.9 liter/min 이었으나 연장파이프가 있을 때는 2.3 liter/min으로 나타났다. 이러한 현상에서 양수하는 물의 온도가 낮고 유량이 부족한 경우에는 연장파이프를 이용하는 것이 좋고, 냉각수치 양은 풍부하지만 그 온도가 낮지 않은 경우에는 연장파이프를 이용하지 않는 것이 좋다는 사실을 알 수 있다. 실험에서의 응축기내 냉각수의 최대 열교환량은 95.75 kJ/min로 나타났다. 작동물질가열탱크와 기액 분리탱크 내의 압력은 0.13­0.14 MPa.a, 물탱크와 응축기내의 압력은 약 0.11 MPa.a정도로 나타났다.

• Asian Pacific Journal of Cancer Prevention
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• 제16권1호
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• pp.125-131
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• 2015

Currently, taxol is mainly extracted from the bark of yews; however, this method can not meet its increasing demand on the market because yews grow very slowly and are a rare and endangered species belonging to first-level conservation plants. Recently, increasing efforts have been made to develop alternative means of taxol production; microbe fermentation would be a very promising method to increase the production scale of taxol. To determine the activities of the taxol extracted from endophytic fungus N. sylviforme HDFS4-26 in inhibiting the growth and causing the apoptosis of cancer cells, on comparison with the taxol extracted from the bark of yew, we used cellular morphology, cell counting kit (CCK-8) assay, staining (HO33258/PI and Giemsa), DNA agarose gel electrophoresis and flow cytometry (FCM) analyses to determine the apoptosis status of breast cancer MCF-7 cells, cervical cancer HeLa cells and ovarian cancer HO8910 cells. Our results showed that the fungal taxol inhibited the growth of MCF-7, HeLa and HO8910 cells in a dose-and time-dependent manner. IC50 values of fungal taxol for HeLa, MCF-7 and HO8910 cells were $0.1-1.0{\mu}g/ml$, $0.001-0.01{\mu}g/ml$ and $0.01-0.1{\mu}g/ml$, respectively. The fungal taxol induced these tumor cells to undergo apoptosis with typical apoptotic characteristics, including morphological changes for chromatin condensation, chromatin crescent formation, nucleus fragmentation, apoptotic body formation and G2/M cell cycle arrest. The fungal taxol at the $0.01-1.0{\mu}g/ml$ had significant effects of inducing apoptosis between 24-48 h, which was the same as that of taxol extracted from yews. This study offers important information and a new resource for the production of an important anticancer drug by endofungus fermentation.

• Asian Pacific Journal of Cancer Prevention
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• 제13권7호
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• pp.3519-3528
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• 2012

Inhibitory effects of Maclura amboinenesis Bl, one plant used traditionally for the treatment of cancers, on metastatic potential of highly metastatic B16F10 melanoma cells were investigated in vitro. Cell proliferation was assessed using the MTT colorimetric assay. Details of metastatic capabilities including invasion, migration and adhesion of B16F10 melanoma cells were examined by Boyden Chamber invasion and migration, scratch motility and cell attachment assays, respectively. The results demonstrated that n-hexane and chloroform extracts exhibited potent anti-proliferative effects (p<0.01), whereas the methanol and aqueous extracts had less pronounced effects after 24 h exposure. Bioactivity-guided chromatographic fractionation of both active n-hexane and chloroform extracts led to the isolation of two main prenylated xanthones and characterization as macluraxanthone and gerontoxanthone-I, respectively, their structures being identified by comparison with the spectral data. Interestingly, both exhibited potent effective effects. At non-toxic effective doses, n-hexane and chloroform extracts (10 and $30{\mu}g/ml$) as well as macluraxanthone and gerontoxanthone-I (3 and $10{\mu}M$) significantly inhibited B16F10 cell invasion, to a greater extent than $10{\mu}m$ doxorubicin, while reducing migration of cancer cells without cellular cytotoxicity. Moreover, exposure of B16F10 melanoma cells to high concentrations of chloroform ($30{\mu}g/ml$) and geratoxanthone-I ($20{\mu}M$) for 24 h resulted in delayed adhesion and retarded colonization. As insights into mechanisms of action, typical morphological changes of apoptotic cells e.g. membrane blebbing, chromatin condensation, nuclear fragmentation, apoptotic bodies and loss of adhesion as well as cell cycle arrest in the G1 phase with increase of sub-G1 cell proportions, detected by Hoechst 33342 staining and flow cytometry were observed, suggesting DNA damage and subsequent apoptotic cell death. Taken together, our findings indicate for the first time that active n-hexane and chloroform extracts as well as macluraxanthone and gerontoxanthone-I isolated from Maclura amboinensis Bl. roots affect multistep of cancer metastasis processes including proliferation, adhesion, invasion and migration, possibly through induction of apoptosis of highly metastatic B16F10 melanoma cells. Based on these data, M. amboinensis Bl. represents a potential candidate novel chemopreventive and/or chemotherapeutic agent. Additionally, they also support its ethno-medicinal usage for cancer prevention and/or chemotherapy.