Induction of Apoptosis in SNU-16 Human Gastric Cancer Cells by the Chloroform Fraction of an Extract of Dangyuja (Citrus grandis) Leaves

문정용,Hana Kim,조문제,Weon Young Chang,Cheong-Tae Kim,김소미 한국응용생명화학회 2009 Journal of Applied Biological Chemistry (J. Appl. Vol.52 No.2

The anticancer effect of Dangyuja (Citrus grandis Osbeck) leaf extract was investigated using SNU-16 human gastric cancer cells. Maximum cytotoxicity was observed from the chloroform fraction (CF) of the extract. Cell death was dose-dependent (IC50=ca. 92.15 μg/mL), and was characterized by apoptotic body formation and DNA fragmentation. Flowcytometric analysis showed that treatment of CF resulted in a marked accumulation of cells in the sub-G1 phase. The induction of apoptosis was confirmed by caspase-3 activity assays and immunoblotting using antibodies against Bid, Bcl-2, Bax, poly (ADP-ribose) polymerase, caspase-9, caspase-8, caspase-7, and caspase-3. Analyses of the CF by gas chromatography (GC) and GC-mass spectrometry tentatively identified 21 compounds, including linoleic acid, linolenic acid, palmitic acid, α-amyrin, γ-sitosterol, dihydrolanosterol, methyl palmitate, and nobiletin. These results provided the first evidence that the CF of a Dangyuja leaf extract induces apoptosis in SNU-16 cells. Our findings may lead to new strategies for the treatment of human gastric cancer.

인간 유방암 줄기세포에서 레몬잎 메탄올 추출물의 항암 효능

문정용,( Linh Thi Thao Nguyen ),현호봉,( Ahmed Osman ),조민환,한수영,이동선,안광석 한국응용생명화학회 2015 Journal of Applied Biological Chemistry (J. Appl. Vol.58 No.3

본 연구에서는 레몬 잎 메탄올 추출물(MLL)의 인간 유방암 줄기 세포인 MCF-7-SC에 대한 항암 활성을 조사하였다. MLL이MCF-7-SC에서 apoptosis를 유도하였으며, 이를 apoptotic body의 형성, sub-G1 phase 및 annexin V-positive 세포와 Bax/Bcl-2 ratio의 증가, caspase-9과 caspase-3의 활성화 및 PARP의 절단을 통하여 확인하였다. 동시에 MCF-7-SC에서 MLL은 acidic vesicular organelles의 형성, LC3-II의 축적 증가, Akt/mTOR/p70S6K의 활성 억제 등을 통하여 autophagy를 유도하였다. Epithelial-mesenchymal transition (EMT)는 세포가 전이 상태를 획득하기 위한 중요한 과정이며, 이 기작은 암세포가 전이되는 것을 억제함에 있어서 중요한 표적이 된다. 낮은 농도에서의 MLL은 epithelial 마커 단백질인 E-cadherin이 증가와 mesenchymal 마커 단백질인 Snail과 Slug의 발현 감소를 통해 EMT 과정을 저해함으로써 MCF-7-SC에서 항전이 활성을 나타내었다. 본 연구에서는 레몬 잎 메탄올 추출물이 농도 의존적으로 유방암 줄기세포에 대해 세포 독성과 항전이 활성을 나타내고 있으며, 따라서 레몬잎은 항암 소재로서의 개발 가능성이 높은 식물이라고 사료된다. The anticancer activity of a methanolic extract from lemon leaves (MLL) was assessed in MCF-7-SC human breast cancer stem cells. MLL induced apoptosis in MCF-7-SC, as evidenced by increased apoptotic body formation, sub-G1 cell population, annexin V-positive cells, Bax/Bcl-2 ratio, as well as proteolytic activation of caspase-9 and caspase-3, and degradation of poly (ADP-ribose) polymerase (PARP) protein. Concomitantly, MLL induced the formation of acidic vesicular organelles, increased LC3-II accumulation, and reduced the activation of Akt, mTOR, and p70S6K, suggesting that MLL initiates an autophagic progression in MCF-7-SC via the Akt/mTOR pathway. Epithelialmesenchymal transition (EMT), a critical step in the acquisition of the metastatic state, is an attractive target for therapeutic interventions directed against tumor metastasis. At low concentrations, MLL induced anti-metastatic effects on MCF-7-SC by inhibiting the EMT process. Exposure to MLL also led to an increase in the epithelial marker E-cadherin, but decreased protein levels of the mesenchymal markers Snail and Slug. Collectively, this study provides evidence that lemon leaves possess cytotoxicity and antimetastatic properties. Therefore, MLL may prove to be beneficial as a medicinal plant for alternative novel anticancer drugs and nutraceutical products.

류사보험(類似保險)의 업무령역(業務領域) 확대(擴大)에 따른 생보업계(生保業界) 대응방안(對應方案)

문정용 보험연구원 1993 보험금융연구 Vol.9 No.-

Free Radical-scavenging Activities and Cytoprotective Effect of Polyphenol-rich ethyl acetate Fraction of Guava (Psidium cattleianum)Leaves on H2O2-treated HepG2 Cell

문정용,김소미,이사라,정승원,김종찬,안광석,Ashik Mosaddik 한국응용생명화학회 2013 Applied Biological Chemistry (Appl Biol Chem) Vol.56 No.6

Total phenolic contents of different fractions of the Psidium cattleianum leaf extract and their antioxidant capacity against several free radicals were examined. Protective effect of the ethyl acetate fraction (EAF) on H2O2-induced DNA damage in HepG2 cells were also evaluated, and the phytochemical profile of EAF was analyzed using tandem mass spectrometry. EAF derived from the 80% methanol extract of the leaf contained a remarkable amount of polyphenol and showed high levels of DPPH and alkyl radical scavenging activity, promoted cell viability, and protected against H2O2-induced DNA damage in HepG2 cells. Phytochemical analysis revealed that the major components in the EAF included quercetin monoglycoside, phloridizin, quercetin 3-diglycoside,quercetin-3-glucuronide, 2,6-dihydroxy-3,5-dimethyl-4-glucopyranosylbenzophenone,phenolic acid, guaijaverin, and naringin. The present study suggests possible synergistic or competitive antioxidant action of the major compounds of cattley guava leaf on H2O2-induced DNA damage in HepG2 cells. These results indicate that the ethyl acetate fraction of the guava leaf could be used as a potential source of natural antioxidants, and these findings will facilitate the utilization of guava leaf as a source of functional food.

고등학생의 지구과학 영역별 개념 이해의 수준 분석

문정용 ( Jeong-yong Mun ),나건민 ( Geon-min Na ),김학성 ( Hak-sung Kim ),조재희 ( Jae-hee Cho ) 한국교원대학교 과학교육연구소 2018 청람과학교육연구논총 Vol.24 No.1

이 연구의 목적은 고등학생들이 지구과학 분야에서 가지고 있는 개념 이해의 수준을 파악하여 올바른 과학적 개념을 형성할 수 있는 교수학습 방안을 마련하는 데 있다. 이를 위해 고등학교 1, 2학년 학생을 대상으로 고체지구, 대기와 해양, 우주 영역으로 구분하여 각 영역별 개념 이해의 수준을 분석하였다. 교사의 명확하지 않은 설명에 대하여 여과 없이 무의식적으로 받아들이거나 학생의 과학적인 관심이 낮아 과학적 개념과는 다른 오개념을 형성하는 것으로 나타났다. 이미 형성된 오개념에 대하여 적절한 교수 학습 모형을 활용하거나 교수 학습 자료 및 교구 개발을 통하여 올바른 과학적 개념을 형성하도록 도움을 줄 수 있다. This study is to analyze on high school students' levels of understanding concepts in the earth science subject in a small city in South Korea. The earth science subject is composed of diverse concepts in such fields as ‘the solid earth’, ‘the atmosphere and the ocean', and ‘the space'. The students who are the first and second graders in high school held misconceptions in the earth science subject since the students either unconsciously accepted ambiguous explanations of teacher or had a low level of interest in science. It is important to adopt appropriate teaching and learning methods in introducing scientific concepts in the earth science subject so that high school students may not develop misconceptions.

미성숙 감귤 과피 초임계 추출물의 성분 분석과 자궁암세포 성장억제효능

문정용(Moon, Jeong Yong),송연우(Song, YeonWoo),현호봉(Hyun, Ho Bong),김소미(KimCho, Somi) 한국산학기술학회 2015 한국산학기술학회논문지 Vol.16 No.12

본 연구의 목적은 제주도 재래감귤인 팔삭과 이예감 초임계 추출물에 대한 인간 자궁암 세포인 HeLa 세포에서의 성장억제 효능을 탐색하고, 각 추출물에서의 유효 활성 성분을 분석하는데 있다. HeLa 세포에서의 성장억제 및 세포사멸 효능을 탐색하기 위해 MTT assay와 Hoechst 33342 염색을 수행하였으며, 성분분석은 가스크로마토그래피 질량분석기 (GC/MS)를 이용하였다. 두 종류의 미성숙 감귤 과육, 과피 초임계 추출물에 대한 HeLa 세포의 성장 억제 효능을 비교해 본 결과, 과피 추출물은 팔삭과 이예감 모두 비슷한 세포사멸 효능을 나타냈다. 과피 추출물을 처리한 세포에서는 농도 의존 적으로 성장률이 억제되었으며, 처리 농도 400 μg/mL에서 팔삭과 이예감 추출물이 각각 87.16%와 92.95%의 세포사멸 효능 을 나타냈으나, 과육 추출물의 세포 성장 억제 효능은 처리 농도 400 μg/mL까지 관찰되지 않았다. Hoechst 33342 염색을 통해 apoptotic body 형성을 현미경으로 관찰한 결과, 100, 200 μg/mL 과육 초임계 추출물을 처리한 세포에서는 apoptpotic body를 관찰할 수 없었으나, 동일한 농도의 과피 추출물을 처리한 세포의 경우에서는 apoptotic body가 현저하게 증가하는 것을 확인 할 수 있었다. GC/MS 분석을 통해 미성숙 팔삭 과육과 과피 초임계 추출물에서 각각 27개와 31개의 화합물을 검출하였으며, 미성숙 이예감 과육과 과피 초임계 추출물에서는 각각 27개와 29개의 화합물을 검출하였다. 팔삭 과피 초임계 추출물에는 1,1,4,4-Tetramethyl- 2-tetralone(20.86%), alloimperatorin(8.15%), limonene (11.23%), auraptene(7.29%) 등이 주로 함유되어 있었으며, 이예감 과피 초임계 추출물에는 limonene(22.19%), linalool(11.23%), γ-sitosterol(9.12%) 등이 주로 함유되 어 있었다. This study was performed to investigate the antiproliferative activities of supercritical extracts from phalsak(Citrus hassaku Hort ex Tanaka) and yeagam(Citrus iyo Hort. ex Tanaka) against human cervical carcinoma HeLa cells and the chemical compositions of the extracts. The anticancer properties of supercritical extracts were demonstrated using the MTT assay and Hoechst 33342 staining and the compositional analyses were conducted by using gas chromatography-mass spectrometry(GC-MS). The peel extracts of both species exhibited similar antiproliferative effect. The antiproliferative activity of the flesh extracts was not detected up to 400 μg/mL, whereas peel extracts of phalsak and yeagam reduced cell viability with 87.16% and 92.95% at 400 μg/mL, respectively. There was a dramatic increase of the apoptotic body formation in the cell treated with peel extracts while no apoptotic body formation detected in the cell treated with flesh extracts at 100, 200 μg/mL. By GC-MS analysis, 27 and 31 kinds of compounds identified in flesh and peel of phalsak, while 27 and 29 kinds of compounds were identified in flesh and peel of yeagam, respectively. 1,1,4,4-Tetramethyl- 2-tetralone(20.86%), alloimperatorin(8.15%), limonene(11.23%), and auraptene(7.29%) were major in peel of phalsak, whereas limonene(22.19%), linalool(11.23%), and γ -sitosterol(9.12%) were major in peel of yeagam. som

유산균 발효에 의한 골드키위 발효물의 항산화 효능 증진

류지연,박현정,문정용,김찬식,김소미 한국식품저장유통학회 2018 한국식품저장유통학회지 Vol.25 No.2

Gold kiwifruit was fermented with Lactobacillus plantarum CK10 derived from kimchi and the fermented products were extracted with ethanol at various fermentation time-points The bacterial cellular density, total titratable acidity, total polyphenol content (TPC), and total flavonoid content (TFC) increased during fermentation, while pH values and total soluble solids decreased. Levels of TPC and TFC were highest after five days, at 1.21±0.13 mg GAE/g dry weight and 0.36±0.04 mg RE/g dry weight, respectively. The antioxidant activities of the fermented gold kiwifruit were analyzed using Fe2+ chelating activity, 2,2-diphenyl-1-picrylhydrazyl (DPPH)-, and 2,2'-azino-bis(3- ethylbenzothiazoline-6-sulphonic acid) (ABTS)- radical scavenging activities, and superoxide dismutase (SOD)-like activity. The Fe2+ chelating activity of gold kiwifruit (125-500 μg/mL) peaked after five days of fermentation at 38.40-78.47%. The DPPH radical- scavenging activity and SOD-like activity were somewhat higher after seven days of fermentation (36.01-86.81% and 54.79-93.83% at 2.5-10.0 mg/mL concentration of samples, respectively). On the other hand, the ABTS radical- scavenging activity of fermented gold kiwifruit was similar to that of the non-fermented form. The polyphenol and flavonoid contents were significantly correlated with the antioxidant activity. In conclusion, our results suggest that TPC, TFC, and antioxidant activity were increased after five and seven days of fermentation, respectively. Therefore, fermented gold kiwifruit with its increased antioxidant activity could be useful in the development of functional foods.

녹차와 로즈마리 추출물을 첨가한 두유의 항산화 활성

류지연,송연우,문정용,전능재,김소미 한국식품저장유통학회 2017 한국식품저장유통학회지 Vol. No.

The objective of this study was to compare antioxidant activities of green tea, rosemary, lemon and bamboo leaves extracts using three different extraction methods (80% ethanol sonication extraction, distilled water autoclave extraction, distilled water extraction at room temperature). As a result, green tea and rosemary extracts showed relatively high antioxidant activities compared with those of lemon and bamboo leaves. These green tea and rosemary extracts were subsequently added to soymilk and their mixtures were examined for antioxidant activities. Soymilk added with 10% green tea distilled water autoclave extracts and distilled water at room temperature extracts showed relatively high DPPH, 85.09±2.26%, 84.38±1.97%, and ABTS radical scavenging activities, 73.43±2.78%, 81.34±4.78%, respectively. On the other hand, soymilk added with 10% rosemary distilled water autoclave extracts showed remarkably higher DPPH radical scavenging activity while their ABTS radical scavenging ability was similar to that of non-added soymilk. In conclusion, our results suggest that the addition of green tea distilled water autoclave extracts, distilled water at room temperature extracts and rosemary distilled water autoclave extracts to soymilk could contribute to the development of added value soymilk products with increased antioxidant activities.

녹차와 로즈마리 추출물을 첨가한 두유의 항산화 활성

류지연,송연우,문정용,전능재,김소미,Ryu, Ji-yeon,Song, YeonWoo,Moon, Jeong Yong,Jun, Neung-Jae,Kim Cho, Somi 한국식품저장유통학회 2017 한국식품저장유통학회지 Vol.24 No.6

제주산 녹차, 로즈마리, 레몬 잎, 조릿대 잎을 물과 에탄올을 이용해 다양한 조건에서 추출물을 제조하여 각 추출물에 대한 항산화 활성을 측정하였다. 그 결과, 녹차와 로즈마리 추출물이 레몬과 조릿대 잎 추출물과 비교하여 상대적으로 높은 항산화 활성을 보였다. 항산화 활성이 상대적으로 우수한 녹차와 로즈마리 추출물들을 두유에 첨가한 뒤, 혼합물의 항산화 활성을 측정하였다. 녹차 증류수 가압 가열 추출물과 상온 정치 추출물 10% 첨가한 두유에서 상대적으로 가장 높은 활성을 보였으며, 녹차 증류수 가압 가열 추출물과 상온 정치 추출물의 DPPH 라디칼 소거능은 각각 $85.1{\pm}2.3%$, $84.4{\pm}2.0%$을, ABTS 라디칼 소거능은 각각 $73.4{\pm}2.8%$, $81.3{\pm}4.8%$로 확인되었다. 한편, 로즈마리 증류수 가압 가열 추출물 10% 첨가한 두유에서 무첨가 두유에 비해 더 높은 DPPH 라디칼 소거능이 확인되었으며, ABTS 라디칼 소거능은 무첨가 두유와 유사했다. 따라서 녹차 증류수 가압 가열추출물과 상온 정치 추출물, 그리고 로즈마리 증류수 가압 가열 추출물을 두유에 첨가 시 항산화 활성이 향상된 고부가가치 두유제품 개발 제발이 가능할 것으로 기대된다. The objective of this study was to compare antioxidant activities of green tea, rosemary, lemon and bamboo leaves extracts using three different extraction methods (80% ethanol sonication extraction, distilled water autoclave extraction, distilled water extraction at room temperature). As a result, green tea and rosemary extracts showed relatively high antioxidant activities compared with those of lemon and bamboo leaves. These green tea and rosemary extracts were subsequently added to soymilk and their mixtures were examined for antioxidant activities. Soymilk added with 10% green tea distilled water autoclave extracts and distilled water at room temperature extracts showed relatively high DPPH, $85.09{\pm}2.26%$, $84.38{\pm}1.97%$, and ABTS radical scavenging activities, $73.43{\pm}2.78%$, $81.34{\pm}4.78%$, respectively. On the other hand, soymilk added with 10% rosemary distilled water autoclave extracts showed remarkably higher DPPH radical scavenging activity while their ABTS radical scavenging ability was similar to that of non-added soymilk. In conclusion, our results suggest that the addition of green tea distilled water autoclave extracts, distilled water at room temperature extracts and rosemary distilled water autoclave extracts to soymilk could contribute to the development of added value soymilk products with increased antioxidant activities.

Mango (Mangifera indica L.) Peel Extracts Inhibit Proliferation of HeLa Human Cervical Carcinoma Cell via Induction of Apoptosis

Md. Ramjan Ali,문정용,Rajendra Gyawali,Ashik Mosaddik,Youn-Chul Ryu,김소미 한국응용생명화학회 2012 Applied Biological Chemistry (Appl Biol Chem) Vol.55 No.3

Flesh and peel extracts of five mango varieties were compared for their antioxidant and antiproliferative activities. Fozli peel (FP) extract was found to have the highest antioxidant activity and to significantly inhibit the proliferation of HeLa human cervical carcinoma cells in a dose-dependent manner. Hoechst 33342 staining revealed apoptotic nuclei with condensed chromatin, and cell growth arrest was confirmed by flow cytometry. Apoptotic signaling induced by FP extract was characterized by an increased Bax/Bcl-2 ratio and down-regulation of Bcl-2. Moreover, FP extract treatment triggered the proteolytic activation of caspases-3, -8, and -9 and the degradation of poly ADP-ribose polymerase in HeLa cells. These results indicate that FP is an excellent source of phenolic and/or flavonoid compounds, and may have applications in the treatment of human cervical carcinoma and as an antioxidant in the food and pharmaceutical industries. Gas chromatography-mass spectrometry analysis of FP extract revealed that 2,5-dihydroxyphenol was detected with highest amount and other compounds such as 5-hydroxymethyl-2-furancarboxaldehyde, pentadecanoic acid, (Z,Z)-9,12-octadecadienoic acid, and 2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one were also found in high amounts.