• 생명과학회지
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• 제22권1호
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• pp.41-48
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• 2012

봉독은 동양의학에서 관절염, 류마티즘 및 각종 암을 포함하여 다양한 질병을 치료하기 위하여 이용되었다. 최근 봉독의 신생혈관 억제효과에 대한 연구가 보고되었으나 정확한 분자메커니즘에 대해서는 보고가 미흡하다. 따라서, 본 연구는 봉독이 인간결장암세포인 HCT116세포에서 신생혈관생성과 종양진행에 중요한 역할을 하는 HIF-$1{\alpha}$와 VEGF 발현 억제효과를 조사하였다. 그 결과 봉독은 $CoCl_2$로 유도한 저산소 상태에서 VEGF와, HIF-$1{\alpha}$의 발현을 감소시키며 HIF-$1{\alpha}$의 promoter 영역인 HRE 활성을 억제하였다. 이러한 봉독의 HIF-$1{\alpha}$ 발현억제효과는 ERK1/2의 인산화 조절을 통한 것이며, 봉독은 p38, JNK, AKT의 인산화에는 영향을 끼치지 않았다. 또한 봉독의 효과를 나타내는 단일물질 탐색을 위해 봉독의 생리활성 물질로 알려진 아파민과 멜리틴을 조사한 결과, HIF-$1{\alpha}$와 VEGF 억제효과는 아파민에 기인하는 것이라고 예상 할 수 있었다. 이와 같은 결과를 통하여 본 연구에서는 봉독의 혈관신생 억제에 대한 새로운 신호전달기전 및 인간 결장암세포 전이 억제제로서의 잠재성을 확인하였다.

• 생명과학회지
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• 제21권11호
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• pp.1518-1525
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• 2011

쏘라페닙은 간암 치료제로 승인된 유일한 약이다. 전세계 암환자들의 인삼추출물 사용이 증가 되고 있지만 쏘라페닙과의 상호작용에 대한 연구는 부족하다. 사람의 간암 세포주와 생쥐 모델을 사용하여 쏘라페닙과 인삼추출물의 약물 상호작용을 알아보고자 하였다. 저농도 인삼추출물 투여시 암세포주의 성장과 pERK(phosphorylation of extracellular signal-regulated kinase)의 증가가 관찰되었고 고농도 투여시 암세포 억제와 pERK 감소가 관찰되었다. 성장 사이클이 없는 세포에서 쏘라페닙의 항암 효과가 감소한 반면 저농도 인삼 투여 시 항암 효능이 증진되어 나타났다. PD98059 (ERK 인산화 억제재)은 효과적으로 ERK 인산화를 억제하여 인삼추출물의 쏘라페닙 감작 작용을 억제시켰다. 생쥐 간암 세포주 모델에서, 저농도 인삼추출물은 다소 암세포 크기를 증가 시켰지만 고농도 투여시 감소시켰다. 그러나, 인삼추출물과 쏘라페닙 동시 투여시 항암 효능은 현저히 증가되었다. 정상조직에서 저농도 인삼에 의해 PERK 증가가 관찰되었으며 이것은 홍삼에 의한 독성 증가와 관련될 것으로 추정되었다. 결론적으로 인삼추출물과 쏘라페닙은 농도에 따라 항암효능을 증가 시킬 수 있음을 보여 주었지만 독성의 가능성도 함께 증가시켰다. 인삼추출물과 쏘라페닙 약물 상호작용에 대한 더 면밀한 연구가 필요할 것으로 보인다.

• 생명과학회지
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• 제18권1호
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• pp.120-128
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• 2008

본 연구에서는 지금초 열수 추출물(WEEH)의 처리에 의한 TRAIL 저항성 인체 위암세포의 증식 억제와 연관된 apoptosis유발에 관한 기전 해석을 시도하였다. 이를 위하여 AGS 세포주가 사용되었으며 다음과 같은 결과를 얻었다. 본 연구에서 조사된 범위 내에서의 TRAIL (200 ng/ml) 및 WEEH (0.04 mg/ml)의 단독 처리에 의한 AGS 세포에 유의적인 세포 독성을 나타내지 않았다. 그러나 TRAIL 및 WEEH의 혼합 처리는 WEEH의 처리 농도의존적으로 AGS 세포의 증식을 억제하였으며, 이는 apoptosis 유발에 의한 것임을 MTT assay, 핵의 염색질 응축 및 세포주기 sub-G1기에 속하는 세포빈도의 증가 등으로 확인하였다. TRAIL 및 WEEH 혼합 처리에 의한 apoptosis 유발 기전의 해석을 위하여 다양한 생화학적 분석법에 의하여 조사된 결과에 의하면, TRAIL 및 WEEH 혼합처리에 의한 caspase-8의 활성화에 의한 BID의 truncation화 및 이와 연관된 미토콘드리아 기능의 손상에 따른 caspase-9의 활성화와 연관성이 있었다. 이러한 미토콘드리아 기능 손상 및 caspase-9의 활성화는 Bcl-2, Bcl-xL, XIAP 및 cIAP-2등과 같은 anti-apoptotic 인자들의 발현 저하와 연관성이 있는 것이며, 이로 인한 caspase-3의 활성화에 의한 PARP 단백질의 단편화 유도로 apoptosis가 일어난 것으로 예측되어 진다. 비록 부가적인 연구들이 요구되어지지만, 본 연구의 결과는 TRAIL저항성 암세포의 항암전략에 지금초 추출물의 적용 가능성을 보여주는 것으로서 지금초의 항암작용의 규명에 중요한 자료를 제공하여 줄 것으로 생각된다.

• 대한한방부인과학회지
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• 제26권1호
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• pp.25-40
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• 2013

Objectives: The object of this study was to observe the in vitro antibacterial effects of five single(Pulsatillae Radix, Patrinae Radix, Sanguisorbae Radix, Sophorae Flos, and Sophorae Radix) aqueous herbal extracts, traditionally used for treating various gynecological diseases including mastitis in Korea, against Staphylococcus aureus. Methods: Antibacterial activities against Staphylococcus aureus of aqueous extracts of Pulsatillae Radix, PatrinaeRadix, Sanguisorbae Radix, Sophorae Flos, and Sophorae Radix were detected using standard agar microdilution methods. In addition, the effects on the bacterial growth curve were also monitored at Minimal Incubation Concentration(MIC) and $MIC{\times}2$ levels. The effects on the intracellular killing and bacterial invasion of individual test materials were also observed using murine macrophage(Raw 264.7) and human mammary gland carcinoma cell(MCF-7). Results: MIC of aqueous extracts of Pulsatillae Radix, Patrinae Radix, Sanguisorbae Radix, Sophorae Flos, and Sophorae Radix against Staphylococcus aureus were detected as $0.215{\pm}0.107$ mg/ml, $0.273{\pm}0.107$ mg/ml, $0.469{\pm}0.297$ mg/ml, $11.850{\pm}8.406$ mg/ml, and $0.664{\pm}0.546$ mg/ml, respectively. MIC of Ciprofloxacin was detected as $0.469{\pm}0.297{\mu}g/ml$ at same conditions. In addition, all five single aqueous herbal extracts were also showed marked dosage-dependent inhibition of bacterial growth. The effects of intracellular killing with Raw 264.7 and inhibition of bacterial invasion with MCF-7 cells were detected, in the order of Sophorae Flos, Pulsatillae Radix, Patrinae Radix, Sanguisorbae Radix and Sophorae Radix aqueous extracts in the present study. Conclusions: The results obtained in this study suggest that all five single aqueous herbal extracts showed antibacterial effects against Staphylococcus aureus and they also showed dosage-dependent inhibitory effects on the bacterial growth. They showed the significant intracellular killing and inhibition of bacterial invasion effects. It means, all five single aqueous herbal extracts may show potent anti-infectious effects against Staphylococcus aureus for mastitis.

• 한국식품영양과학회지
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• 제40권12호
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• pp.1694-1699
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• 2011

B. subtilis-SKm, B. subtilis HJ18-4, B. subtilis KCCM 42923을 스타터로 이용하여 $40^{\circ}C$에서 72시간 발효시켜 청국장(BS-c, BH-c, BK-c)을 제조하였으며, 이때 자연 발효하여 제조한 청국장(NF-c)과 함께 발효 특성을 비교, 관찰하였다. pH는 BS-c, BK-c 및 NF-c에서 7.6으로 비슷하게 나타났고, BH-c가 5.9로 가장 낮았다. 호기성 균수는 BS-c, BK-c, NF-c에서 10.0 log cfu/g대로 비슷한 분포를 보였고, BH-c의 경우는 9.7 log cfu/g로 비교적 낮게 나타났다. 아미노태 및 암모니아태 질소 함량과 ${\gamma}$-GTP의 역가의 결과를 관찰한 결과, 모두 BS-c에서 높게 나타나 발효가 가장 잘 이루어졌음을 알 수 있었으며, BH-c의 경우 발효 정도가 낮음을 알 수 있었다. Protease와 ${\alpha}$-amylase 활성 역시 BS-c에서 높게 나타났다. 관능평가에서는 4종의 청국장이 대체로 비슷한 기호도를 나타냈으나, 종합적인 맛과 종합적인 평가에서 BS-c가 가장 높은 선호도를 나타내었다. 또한 DPPH free radical 소거 효과와 HT-29 암세포 성장 억제효과는 스타터를 이용하여 제조된 청국장인 BS-c, BH-c, BK-c가 NF-c에 비해 우수한 효과를 보였으며, 역시 BS-c가 가장 높은 효과를 나타냈다. 따라서 스타터의 종류에 따라 청국장의 품질 및 기능성에 차이가 나타나며, B. subtilis-SKm을 스타터로 이용할 경우 청국장의 품질 및 기능성 증진 효과를 기대할 수 있다.

• 대한두경부종양학회지
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• 제22권1호
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• pp.8-14
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• 2006

Objectives: The p53 tumor suppressor gene has a key role in cellular control mechanisms involving apoptosis and DNA repair, leading to the G1 arrest following DNA damage. Its mutation is one of the most frequent alterations in human cancers. Ki-67 is identified in replicating cells of both benign and malignant lesions, so it can be the predictor of proliferative activity. The aim of this study is to evaluate the expression of p53 and Ki-67 in salivary gland tumors. Materials and Methods: Immunohistochemical analysis was used to detect expression of p53 and Ki-67 in paraffin-embedded samples from 31 benign and 27 malignant salivary gland tumors. Results were analyzed between benign and malignant tumors and compared with the clinical parameters such as stage and recurrence in malignant tumors. Results: p53 overexpression was detected in 19.6% of benign tumors and 40.7% of malignant tumors, but there was no statistical significance. p53 was significantly expressed in Warthin's tumor(45.5%) compared with pleomorphic adenoma(5.9%). Only 5.9% of pleomorphic adenoma were positive for p53, while 60% of carcinoma ex pleomorphic adenoma were positive for p53. Ki-67 was expressed in 3.2% of benign tumors and 51.9% of malignant tumors, which showed significant higher expression in malignant tumors. In malignant tumors, p53 and Ki-67 expressions bore no correlation to stage and recurrence. Conclusion: p53 overexpression is not associated with the progression of malignant tumors, and Ki-67 overexpression can be used as biologic indicator of malignant salivary gland tumors.

• 대한한의학회지
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• 제27권3호
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• pp.107-131
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• 2006

Background and Aims: Dansamtongmek-tang (DSTMT) and Dansamsengmek-san (DSSMS) have been used for many years as therapeutic agents for the acute stage of cerebrovascular disease, hypertension and hyperlipidemia in Oriental medicine, but the effects of DSTMT and DSSMS on hyperlipidemia and safety for cell damage are not yet well-known. This study was done to investigate the effects of DSTMT and DSSMS on hyperlipidemia. Methods: In vivo test: after administering DSTMT and DSSMS to SHR and ICR occurred hyperlipidemia for 3 weeks, we analyzed body weight, cholesterol levels. TG, HDL-chol, LDL-chol, LDH in plasma, brain, liver and kidney tissue, and DNA by RT-PCR. In vitro test: after administering DSTMT and DSSMS to human hepatocellular carcinoma in hypoxia, we observed cell cohesion by light microscope, analyzed the inflow of Ca2+ by confocal laser scanning microscope and DNA by RT-PCR. Results: DSTMT significantly decreased the levels of triglyceride and increased the levels of HDL-cholesterol in SHR, and significantly decreased the levels of LDL-cholesterol and body weight and increased the levels of HDL-cholesterol in ICR. DSSMS significantly decreased body weight, total cholesterol levels, LDL-cholesterol, LDH and cardiac risk factor (CRE) in SHR and significantly decreased the levels of total cholesterol, triglyceride, LDL-cholesterol, LDH and CRF in ICR. DSTMT had an effect on protecting cells from damage by inhibiting production of p53 mRNA, and in DSSMS, by inhibiting production of p53 mRNA and p21 mRNA after hypoxia. DSTMT effectively blocked off Ca2+ at low density, but DSSMS effectively blocked off Ca2+ at high density. Both DSTMT and DSSMS had an effect on inhibiting lipid metabolism by blocking off production of apo B mRNA. Conclusions: These results suggest that DSTMT and DSSMS might be usefully applied for treatment of hyperlipidemia and suppression of brain damage.

• Asian Pacific Journal of Cancer Prevention
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• 제15권15호
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• pp.6397-6403
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• 2014

Background: Aberrant promoter hypermethylation has been recognized in human breast carcinogenesis as a frequent molecular alteration associated with the loss of expression of a number of key regulatory genes and may serve as a biomarker. The E-cadherin gene (CDH1), mapping at chromosome 16q22, is an intercellular adhesion molecule in epithelial cells, which plays an important role in establishing and maintaining intercellular connections. The aim of our study was to assess the methylation pattern of CDH1 and to correlate it with the expression of E-cadherin, clinicopathological parameters and hormone receptor status in breast cancer patients of Kashmir. Materials and Methods: Methylation specific PCR (MSP) was used to determine the methylation status of CDH1 in 128 invasive ductal carcinomas (IDCs) paired with the corresponding normal tissue samples. Immunohistochemistry was used to study the expression of E-cadherin, ER and PR. Results: CDH1 hypermethylation was detected in 57.8% of cases and 14.8% of normal adjacent controls. Reduced levels of E-cadherin protein were observed in 71.9% of our samples. Loss of E-cadherin expression was significantly associated with the CDH1 promoter region methylation (p<0.05, OR=3.48, CI: 1.55-7.79). Hypermethylation of CDH1 was significantly associated with age at diagnosis (p=0.030), tumor size (p=0.008), tumor grade (p=0.024) and rate of node positivity or metastasis (p=0.043). Conclusions: Our preliminary findings suggest that abnormal CDH1 methylation occurs in high frequencies in infiltrating breast cancers associated with a decrease in E-cadherin expression. We found significant differences in tumor-related CDH1 gene methylation patterns relevant to tumor grade, tumor size, nodal involvement and age at diagnosis of breast tumors, which could be extended in future to provide diagnostic and prognostic information.

• 대한한의학회지
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• 제31권3호
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• pp.34-46
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• 2010

Objective: This experimental study was performed to examine if Hang-Am-Dan non-boiled water extracts (HAD-N) induce apoptosis in human lung carcinoma NCI-H460 cells in vitro and inhibits the growth of NCI-H460 cell-transplanted solid tumor in vivo. Materials and Methods: We cultured NCI-H460 cell lines and xenografted them to nude mice. The mice were divided into 3 groups, NCI-H460 cell alone, NCI-H460 + 90 mg/kg HAD-N treated group, and NCI-H460 + 180 mg/kg HAD-N treated group, with seven mice per group. HAD-N was orally administrated every day for four weeks. We checked their body weight and tumor weight and volumes two times a week and their absolute organ weight and biochemical blood analysis at the final day by sacrificing them. We also calculated their tumor inhibition rate (IR), mean survival time and percent increase in life span (% ILS). Results: In this study, we observed that all of the HAD-N treated mice got smaller tumors. The more doses of HAD-N used, the less IR showed at the 8th day after starting this experiment. Tumor weight and volume of HAD-N treatment groups also decreased. Mean survival time and percent increase in life span (% ILS) in the high-dose HAD-N treatment groups were higher than those of other groups. The test substances in the blood level UN results showed reduction in the significance in both HAD-N 90 mg/kg and HAD-N 180 mg/kg (p<0.01). The blood level phosphatase results in HAD-N 90 mg/kg group compared to NCI-H460 cell alone group showed a reduction in significance (p<0.05). AST levels HAD-N 180 mg/kg group compared to NCI-H460 cell alone group significance as well (p<0.05). Conclusion: We suggest that the results of the in vivo study showed that HAD-N may have potential as a growth inhibitor of tumor-induced NCI-H460 of nude mice in spite of the shortcomings of this study. More studies to overcome those shortcomings and to find out significant antitumor mechanism will be needed.

• 생약학회지
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• 제24권4호
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• pp.267-281
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• 1993

To find antitumor components from higher fungi, the mycelia of Collybia confluens (Pers. ex Fr.) Kummer were cultured in artificial media. For efficient production of the mycelia, the influences of various modifications of culture conditions were examined. A water-soluble protein-bound polysaccharide fraction, Fr. A, was obtained from the mycelia by hot water extraction. When Fr. A was purified and fractionated by DEAE-cellulose and Sepbadex G-200 gel filtration chromatographies into four fractions which were designated B, C, C-I and C-II. The tumor inhibition ratios of these fractions ranged from 46% to 75% against the solid forms of sarcoma 180 in ICR mice at doses of 20 and 50 mg/kg/day when given intraperitoneally. Especially, Fr. C which was named Collyban(CB) exhibited a marked life-prolonging effect of the mice against ascitic forms of sarcoma 180 at a dose of 50 mg via i.p. administration. To extend spectra of the antitumor activities and eliminate the effects of allograft rejection, the characterization of antitumor effects of CB was performed in syngeneic host-tumor systems. It did not show any antitumor activity against L1210 murine leukemia in $CD_2Fl$ mice but prolonged their life span against ascitic forms of $MM_{46}$ carcinoma in $C_3H/He$ mice. Also it exhibited antitumor activity against human cervical cancer HeLa cells that were xenografted into nude mice having BALB/c genetic backgrounds by the i.p. injection at a dose of 100 mg/kg/day. In order to characterize the antitumor components, CB was examined by chemical analysis. It was acidic protein-bound polysaccharides composed of 31% polysaccharide, 27% protein and 3% hexosamine. CB was fractionated into two fractions, Fr. C-I(M.W.: 500 Kd) and Fr. C-II(M.W.:30 and 8 Kd) by Sephadex G-200 gel filtration chromatography.