창이자 및 꿀풀하고초에 의한 NAD(P)H:quinone reductase와 glutathione S-transferase의 유도

손윤희,이기택,박신화,조경희,임종국,남경수,Shon, Yun-Hee,Lee, Ki-Taek,Park, Sin-Hwa,Cho, Kyoung-Hee,Lim, Jong-Kook,Nam, Kyung-Soo 한국생약학회 2001 생약학회지 Vol.32 No.4

Ethanol extracts from Xanthii Fructus (XFE) and Prunellae Spica (PSE) were investigated for the effects on the induction of cancer chemoprevention-associated enzymes. The following effects were measured: (a) induction of quinone reductase (QR) (b) induction of glutathione S-transferase (GST) (c) reduced glutathione (GSH) level. XFE and PSE were potent inducers of quinone reductase activity in Hepa1c1c7 murine hepatoma cells. Glutathione levels were increased with XFE and PSE. In addition, glutathione S-transferase activity was increased with XFE. However, GST activity was not increased with PSE. These results suggest that XFE and PSE have chemopreventive potentials by inducing quinone reductase and increasing GSH levels.

군소내장 분획물의 in vitro에서의 암세포 성장억제 및 quinone reductase 유도 활성 증가 효과

Mi-Ok Shin(신미옥) 한국생명과학회 2010 생명과학회지 Vol.20 No.6

본 연구에서는 군소내장을 각 용매별로 분획하여 암세포 성장 억제 효과와 quinone reductase (QR) 유도 활성증가 효과를 알아보았다. 간암 세포인 HepG2, 유방암 세포주인 MCF-7, 대장암 세포주인 HT-29 그리고 피부암 세포주인 B16-F10 를 이용하여 암세포 성장 억제 효과를 실험한 결과 모든 세포주에서 AKMM층에서 농도 의존적으로 가장 높은 암세포 성장 억제 효과를 나타내었다. 그리고 다음으로 AKMB층, AKMH층 및 AKMA층의 순이었다. 그리고 4종의 암세포주 중에서 B16-F10 세포주가 가장 높은 암세포 성장 억제 효과를 나타내어 특히 피부암에 대한 예방효과가 기대되어진다. 또한 암 예방 효과를 알아보기 위하여 4종의 암세포 중 유일하게 quinone reductase를 가지고 있는 인체 간암세포주인 HepG2를 이용하여 QR 유도 활성 증가 효과를 측정한 결과, 암세포 성장 억제 효과에 있어서 가장 높은 효과를 나타낸 AKMM층에서 가장 높은 QR 유도 활성 증가 효과를 나타내었다. 그러나 다른 분획층에서는 QR 유도 활성 증가 효과를 거의 볼 수 없었다. 이상으로 암세포 성장 억제 효과와 QR 유도 활성 효과에서 모두 methanol 분획층인 AKMM층에서 가장 높게 나타났으므로 이 분획층에 유효한 생리활성 물질이 함유되어 있을 가능성이 추정되어진다. 따라서 폐기되어지는 군소부산물인 내장을 이용한 암 예방 관련 기능성 식품의 개발 가능성이 기대되어지며, 이를 위한 AKMM분획층에 대한 더욱더 심도 높은 집중적인 연구가 요구되어진다. We investigated the growth inhibitory effect of internal organs of Aplysia kurodai (AK) on proliferation in cancer cell lines in vitro. The internal organs of AK were extracted with methanol (AKM), which were then further fractionated into four subfractions by using solvent partition method, resulting in hexane (AKMH), methanol (AKMM), butanol (AKMB), and aqueous (AKMA) soluble fractions. We determined the cytotoxic effect of these four fractions in four kinds of cancer cell lines - HepG2, MCF-7, HT29 and B16-F10 - by MTT assay. Among the four subfractions of AKM, AKMM showed the strongest cytotoxic effects on all cancer cell lines which were used. Morphological changes such as membrane shrinking and blebbing of cells were also observed in AKMM treatment in HepG2 cells. In addition, we also observed quinone reductase (QR) induced effect in the methanol layer (AKMM) of HepG2 cells. AKMM showed the highest induction activity of quinone reductase on HepG2 cells among the partition layers. The QR induced effect of AKMM was determined to be 2.4 at 100 μg/ml level with a control value of 1.0. Although further studies are needed, the present work suggests that internal organs of Aplysia kurodai (AK) may be a chemopreventive agent for the treatment of human cells.

오징어내장 분획물의 in vitro에서의 암세포 성장억제 및 quinone reductase유도 활성 증가 효과

신미옥(Mi-Ok Shin),배송자(Song-Ja Bae) 한국생명과학회 2009 생명과학회지 Vol.19 No.9

본 연구에서는 오징어내장을 각 용매별로 분획하여 암세포 성장 억제 효과와 quinone reductase (QR) 유도 활성 증가효과를 알아보았다. 간암 세포인 HepG2, 유방암 세포주인 MCF-7 그리고 대장암 세포주인 HT-29 를 이용하여 암세포 성장 억제 효과를 실험한 결과 모든 세포주에서 TPMH층에서 가장 높은 암세포 성장 억제 효과를 나타내었고 다음으로 TPMM층에서 도 높은 암세포 성장 억제 효과를 나타내었다. 그리고 HepG2, MCF-7 및 HT-29 세포주에서 모두 농도 의존적인 암세포 성장 억제 효과를 나타내었으나 간암 세포주인 HepG2에서 가장 높은 효과를 나타내어 특히 간암에 대한 예방효과가 기대되어진다. 또한 암 예방 효과를 알아보기 위하여 3종의 암세포 중 유일하게 quinone reductase를 가지고 있는 인체 간암세포주인 HepG2를 이용하여 QR 유도 활성 증가 효과를 측정한 결과, 암세포 성장 억제 효과의 결과에서와는 달리 TPMM층에서 유의적으로 가장 높은 QR 유도 활성 증가 효과를 나타내었고 다음으로 TPMH층에서도 높은 QR 유도 활성 증가 효과를 나타내었다. 본 실험 결과에서 암세포 성장 억제 효과는 오징어내장의 hexane 분획층인 TPMH층에서 가장 높게 나타났으며, QR 유도 활성 효과는 methanol 분획층인 TPMM층에서 가장 높게 나타났으므로 이들 분획층에 유효한 생리활성 물질이 함유되어 있을 가능성 추정되어진다. 따라서 본 연구를 통하여 오징어 부산물인 내장을 이용한 항암 관련 기능성 식품의 개발 가능성이 보이며, 이를 위한 TPMH분획층과 TPMM분획층에 대한 집중적인 연구가 요구된다. We investigated the growth inhibitory effect of internal organs of Todarodes pacificus (TP) on proliferation in human cancer cell lines in vitro. The internal organs of TP were extracted with methanol (TPM), which was then further fractionated into four subfractions by using a solvent partition method, resulting in hexane (TPMH), methanol (TPMM), butanol (TPMB), and aqueous (TPMA) soluble fractions. We determined the cytotoxic effect of these four fractions in three kinds of cancer cell lines - HepG2, MCF-7 and HT-29 - by MTT assay. Among the four subfractions of TPM, TPMH showed the strongest cytotoxic effects at a concentration of 300 μg/ml, displaying 91.56% on the HepG2 cell line and 85.93% on the MCF-7 cell line. Morphological changes such as membrane shirinking and blebbing of cells were also observed during TPMH treatment of HepG2 cells. In addition, we also observed quinone reductase (QR) induced effect in the methanol (TPMM) layer of HepG2 cells. TPMM showed the highest induction activity of quinone reductase on HepG2 cells among the other partition layers. The QR induced effect of TPMM was determined to be 2.7 at a level of 360 μg/ml with a control value of 1.0. Although further studies are needed, the present work suggests that internal organs of Todarodes pacificus (TP) may be a chemopreventive agent for the treatment of human cells.

함초 분획물의 in vitro에서의 암세포 성장억제 및 Quinone Reductase 활성 유도 효과

정복미(Bok-Mi Jung),박정애(Jung-Ae Park),배송자(Song-Ja Bae) 한국식품영양과학회 2008 한국식품영양과학회지 Vol.37 No.2

본 연구에서는 염생식물의 하나인 함초를 각 용매별로 분획하여 암세포 성장억제 효과와 quinone reductase(QR) 활성 유도 효과를 보았다. 간암 세포인 HepG2, 대장암 세포주인 HT-29 그리고 유방암 세포주인 MCF-7을 이용하여 암세포 성장 억제 효과를 실험한 결과 간암세포주인 HepG2에서 제일 효과가 컸고 사용한 3종의 암 세포주 모두 SHMM층에서 월등히 높은 암세포 성장 억제 효과를 나타내었다. 그리고 암 예방 효과를 알아보기 위하여 3종의 암세포 중 유일하게 quinone reductase를 가지고 있는 인체 간암세포주인 HepG2를 이용하여 QR 활성 유도 효과를 측정한 결과 역시 SHMM층에서 유의적으로 QR 활성 유도 효과가 높게 나타났다. 본 실험 결과에서 함초의 methanol 분획층인 SHMM층에서 암세포 성장 억제 효과와 QR 유도 활성 효과가 높게 나타났으며 이 분획층에 유효한 생리활성 물질이 함유되어 있을 가능성이 높을 것으로 추정된다. 본 연구를 통하여 함초를 이용한 암예방 기능성 식품을 개발할 수 있는 가능성이 보이며, 특히 함초의 methanol 분획층에 대한 집중적인 연구가 요구된다. We investigated the growth inhibitory effect of Salicornia herbacea L. (SH) on human cancer cell lines in vitro. SH was extracted with methanol (SHM), followed by further fractionation into four subfractions according to polarity: hexane (SHMH), methanol (SHMM), butanol (SHMB), and aqueous (SHMA) soluble fractions. We determined the growth inhibitory effect of these fractions against human cancer cell lines using MTT assay. Among the four subfractions of SHM, the SHMM showed the strongest cytotoxic effects on cancer cell lines. We also observed quinone reductase (QR)-inducing effect of methanol layer (SHMM) on HepG2 cells and it was determined to be 3.00 at 100 ㎍/mL level compared to the control value of 1.0. The SHMM showed the highest induction activity of quinone reductase on HepG2 cells among the partition layers. The present work suggests that SH merits further study to confirm its chemopreventive potential.

Reduction of Azobenzene by Purified Bovine Liver Quinone Reductase

Kim, Kyungsoon,Shin, Hae-Yong The Korea Science and Technology Center 2000 BMB Reports Vol.33 No.4

Quinone reductase was purified to homogeneity from bovine liver by using ammonium sulfate fractionation, ion-exchange chromatography, and gel filtration chromatography. The enzyme utilized either NADH or NADPH as the electron donor. The enzyme catalyzed the reduction of several quinones and other artificial electron acceptors. Furthermore, the enzyme catalyzed NAD(P)H-dependent reduction of azobenzene. THe apparent Km for 1,4-bezoquinone and azobenzene was 1.64mM and 0.524mM, respectively. The reduction of azobenzene by quinone reductase was almost entirely inhibited by dicumarol or Cibacron blue 3GA, potent inhibitors of the mammalian quinone reductase. In the presence of 1.0μM Cibacron blue 3GA, azoreductase activity was lowered by 45%, and almost complete inhibition was seen above 2.0μM Cibacron blue 3GA.

Reduction of Azobenzene by Purified Bovine Liver Quinone Reductase

Kim, Kyung-Soon,Shin, Hae-Yong Korean Society for Biochemistry and Molecular Biol 2000 Journal of biochemistry and molecular biology Vol.33 No.4

Quinone reductase was purified to homogeneity from bovine liver by using ammonium sulfate fractionation, ionexchange chromatography, and gel filtration chromatography. The enzyme utilized either NADH or NADPH as the electron donor. The enzyme catalyzed the reduction of several quinones and other artificial electron acceptors. Furthermore, the enzyme catalyzed NAD(P)H-dependent reduction of azobenzene. The apparent Km for 1,4-benzoquinone and azobenzene was 1.64 mM and 0.524 mM, respectively. The reduction of azobenzene by quinone reductase was almost entirely inhibited by dicumarol or Cibacron blue 3GA, potent inhibitors of the mammalian quinone reductase. In the presence of 1.0${\mu}M$ Cibacron blue 3GA, azoreductase activity was lowered by 45%, and almost complete inhibition was seen above 2.0 ${\mu}M$ Cibacron blue 3GA.

Induction of Quinone Reductase, an Anticarcinogenic Marker Enzyme, by Extract from Chrysanthemum zawadskii var. latilobum K.

Kim, Ju-Ryoung,Kim, Jung-Hyun,Lim, Hyun-Ae,Jang, Chan-Ho,Kim, Jang-Hoon,Kwon, Chong-Suk,Kim, Young-kyun,Kim, Jong-Sang The Korean Society of Food Science and Nutrition 2005 Preventive Nutrition and Food Science Vol.10 No.4

Induction of NAD(P)H:(quinone-acceptor) oxidoreductase (QR) which promotes obligatory two electron reduction of quinones and prevents their participation in oxidative cycling and thereby the depletion of intracellular glutathione, has been used as a marker for chemopreventive agents. Induction of phase II enzyme is considered to be an important mechanism of cancer prevention. In our previous study, we assessed the quinone reductase QR-inducing activities of 216 kinds of medicinal herb extracts in cultured murine hepatoma cells, BPRc1 and hepalc1c7 cells. Among the 216 herbal extracts tested in that study, extracts from Chrysanthemum zawadskii showed significant induction of QR. In this study, we examined QR-inducing activity of solvent fractions of the herbal extract. The dichloromethane fraction of the herb showed the highest QR induction among the samples fractionated with four kinds of solvents with different polarity. The fraction also significantly induced the activity of glutathione S-transferase (GST), one of the major detoxifying enzymes, at $4{\mu}g/mL\;and\;2{\mu}g/mL$ in hepalc1c7 and BPRc1 cells, respectively. In conclusion, dichloromethane-soluble fraction of Chrysanthemum zawadskii which showed relatively strong induction of detoxifying enzymes merits further study to identify active components and evaluate their potential as cancer preventive agents.

Screening for Korean Vegetables with Anticarcinogenic Enzyme Inducing Activity Using Cell Culture System

Kim, Su-Mi,Ryu, Seung-Hee,Park, Hui-Don,Kim, Sung-Su,Kim, Jeong-Hwan,Kim, Jong-Sang The Korean Society of Food Science and Nutrition 1998 Preventive Nutrition and Food Science Vol.3 No.3

There is extensive evidence suggesting the protective role of fruits and vegetables against chemically induced carcinogenesis. We have tested the ability of a representative range of Korean vegetables to act as blocking agents against neoplastic initiation by determining the induction level of quinone reductase , an anticarcinogenci marker enzyme, in hepalclc 7 cells exposed to vegetable extracts. Among thirty vegetables tested, Arcitum lappa(Burdock), Brassica juncea (Mustard leaf), Pteridium aguilinum (Bracken) and Chrysanthemum cornoratium(Crown daisy) caused a significant induction of quinone teductase activity with a limited increase in arylhdrocarbon hydroxylase activity. Combination of crown daisy with burdock had synergistic effect on quinone reductase induction. Quinone reductase-inducing activity was found mostly in hesane and ehtylactate fractions of MeOH extract of crown daisy while it ws not quinone reductase activity in liver, kideny, lung, and small intestine, confirming the presence of potent QR inducer (s) in crown daisy. These sata suggest that some vegetables including crown daisy induced QR merits further investigation as a potential cancer preventive agent in human.

우뭇가사리 분획물의 항균 및 암세포 성장억제효과

신혜정,강대연,신미옥,배송자,Shin, Hye-Jung,Gang, Dae-Yeon,Shin, Mi-Ok,Bae, Song-Ja 한국해양바이오학회 2007 한국해양바이오학회지 Vol.2 No.2

본 실험은 홍조 해조류의 하나인 우뭇가사리를 추출, 각 용매별로 분획하여 항균효과와 암세포 성장억제 및 QR 유도활성 효과 등의 생리활성을 연구하였다. 우뭇가사리의 각 분획물 GAMM, GAMH, GAMB 및 GAMA층을 단백질 식품 부패 원인균인 Serratia marcescens, Proteus mirabilis의 2종과 감염형 세균성 식중독 원인균인 Salmonella enteritidis, 부패균인 Bacillus subtilis, 식중독 원인균인 Starpylococcus aureus 등 총 5 가지 균종에 처리하여 항균력을 조사한 결과 GAMM층에서 높은 가장 높은 항균 활성 효과를 나타내었다. 또 4종의 인체 암세포주 HT-29, HepG2, MCF-7 및 B16F-10 세포주에 대한 암세포 증식 억제 실험을 한 결과 사용한 4종의 모든 암세포주에서 methanol 분획층인 GAMM에서 낮은 농도의 시료첨가에도 불구하고 괄목할 만한 높은 암세포 성장 억제효과를 나타내었다. 한편, 사용한 4가지 암세포주중 유일하게 quinone reductase를 가지고 있는 HepG2를 이용한 암예 방지표인 quinone reductase 효소 유도 활성 여부를 측정한 결과 분획물 첨가농도를 10, 20, 30 및 $40{\mu}g/mL$로 첨가하였을 때 GAMM의 첨가농도 $20{\mu}g/mL$에서 대조군에 비해 약 2배 이상의 높은 QR유도효과를 나타내었고 최종농도 $40{\mu}g/mL$에서는 2.5배의 암예방 QR 유도효과를 나타내었다. In this study, we investigated antimicrobial and cytotoxicity effects of Gelidium amansii L., which using methanol, dichloromethane and ethanol were extracted and fractionated into four different types : methanol (GAMM), hexane (GAMH), butanol (GAME) and aqueous (GAMA). The antimicrobial activity was increased in proportion to its concentration by the paper disc method. Among the solvent fractions, The methanol partition layer (GAMM) showed the strongest antimicrobial activities and cytotoxic effects on all cancer cell lines. We also observed quinone reductase (QR) induced effects in all fraction layers of GA on HepG2 cells. The QR induced effects of GAMM on HepG2 cells at $40{\mu}g/mL$ concentration indicated 2.5 with a control value of 1.0.

Induction of Quinone Reductase , an Anticarcinogenic Marker Enzyme, by Vitamin E in Both Hepalclc7 Cells and Mice

Kwon, Chong-Suk,Kim, Jong-Sang The Korean Society of Food Science and Nutrition 1999 Preventive Nutrition and Food Science Vol.4 No.2

Induction of NAD(P)H : (quinone-acceptor) oxidoreductase (QR) which obligatory two electron reduction of quinones and prevents their participation in oxidative cycling and thereby the depletion of intracellular glutathione, has been used as a marker for chemopreventive agents. We postulated that vitamin E, an antioxidant, which induces QR as the gene of QR was reported to contain antioxidant reponsive element in the 5'-flanking region. Vitamin E resulted in significant induction of QR in both hepalclc7 cells and mouse tissues. QR induction was observed; to be maximal at 25uM vitamin E for hepalclc7 cells while it was maximal in the level of 2.5∼5 μmoles vitamin E/㎏ BW for mouse tissues. Thus the cancer-preventive effect of vitamin E may be exerted by it induction of intracellular detoxifying enzymes.