약용식물내 에스트로겐성과 항-에스트로겐성을 조사하고 항암인자를 발견하기 위하여, 본연구는 에탄올추출로 제조된 9종류의 한국산 약용식물에 대하여 재조합효모와 MCF-7 사람유방암세포주를 이용하여 스크리닝하고 비교하였다. 재조합효모를 이용한 실험결과, 7종류의 약용식물에서 에스트로겐성이 나타났고, 4종류에서 안드로겐성이 나타났다. 또한 MCF-7 사람유방암세포주를 이용한 실험결과, 8종류의 추출물이 MCF-7 세포의 성장을 억제하는 것으로 확인되었으며 비스페놀 A와 동시 처치한 경우에도 유의적으로 억제하는 것으로 나타났다. 또한 Clyeyrrhiza uralensis, Cassia tora, Syringa velutina, Zingiber officinale, Malva verticillata, Panax ginseng C.A. Meyer는 식물성 에스트로겐으로서 에스트로겐에 양성인 사람유방암세포의 증식을 유의적으로 억제시티는 흥미로운 결과가 제시되었다. 따라서 이번 연구는 한국산 약용식물이 식물성 에스트로겐과 항암인자로서 이용될 수 있으며, 에스트로겐의 활성을 조사하는데 유용하게 이용될 수 있을 것으로 사료된다.

The Sellaginella Tarmariscina is widely used in the traditional oriental herbal medicine for its anti-inflammatory, antioxidant effects. The effects of aqueous extracts of Sellaginella Tarmariscina on the viability and induction of apoptotic cell death were investigated in HL-60 cells. The cell viability after treating with extract of Sellaginella Tarmariscina was quantified by MTT assay. The results showed that could inhibit the proliferation of HL-60 cells and caused a 40% inhibition of HL-60 cells at concentrations of 400 ㎍/ml. The cancericidic effect of Sellaginella Tarmariscina was mediated by apoptosis. Thus, HL-60 cells exposed to Sellaginella Tarmariscina displayed the DNA fragmentation ladder and nucleus chromatin condensation characteristic for apoptosis. In conclusion, our results suggest that the extract of Sellaginella Tarmariscina may induce the apoptotic death in HL-60 cells.

본 연구 결과들을 바탕으로 도깨비부채 잎(RPL)은 GSK3β 활 성화를 통해 IκB-α를 인산화시켜 단백질 분해를 유도하고 Iκ B-α 분해로 인해 p65 핵내 전이를 유도하여 NF-κB 신호전달을 활성화?시킨다. 이러한 NF-κB 신호전달 활성화를 통해 대장암 의 세포생육을 억제하는 것으로 추정된다. 본 결과는 도깨비부채 잎을 소재로 항암을 목적으로 한 천연치료제 및 대체보완소재 개발에 활용할 수 있다고 판단된다. 그러나 도깨비부채 잎의 대장암에 대한 세포생육 억제와 작용기전의 정확한 관련성과 세포생육 억제활성 물질 분석을 위해 추가적인 연구가 필요할 것으로 사료된다.

본 연구에서 상동나무 가지 추출물(STB-E100)은 대장암 세포에서 세포사멸을 유도하여 세포생육을 억제하였다. 또한 Iκ B-α 인산화를 통한 IκB-α 단백질 분해를 유도하며 이로 인해 P65 핵내 전이를 유도하여 NF-κB 신호전달을 활성화시킨다. NF-κB 신호전달 활성화는 GSK3β 활성화를 통해 P65 핵내 전 이를 유도에 의한 것이지만 IκB-α분해는 GSK3β 의존성이 아니다. 상동나무 가지 추출물은 이러한 신호전달 활성화를 통해 세포사멸을 유도하여 대장암의 세포생육을 억제한다. 본 결과를 바탕으로 상동나무 가지가 암 예방 및 치료를 목적으로한 표적 요법에서 항암제 개발의 잠재적 활용 소재로서 이용 가능하다고 사료된다. 그러나 대장암 세포에서 상동나무 가지 추출물에 의해 유도된 NF-κB 신호전달 작용기전을 좀 더 구체적으로 구명할 필요가 있고 대장암에 대한 세포사멸과 작용기전의 정확한 관련성을 조사하기 위해 추가적인 연구가 필요하다.

이상의 연구 결과로 먹넌출 열매 추출물은 GSK3β 의존성 Cyclin D1 단백질의 분해를 통해 대장암세포의 생육 억제와 관련이 있는 것으로 확인된다. 본 결과는 대장암의 항암제 개발을 위한 소재로 먹넌출 열매의 활용이 가능할 것으로 판단된다.

이상의 연구 결과로 미루어 볼 때, 댕댕이나무 잎과 가지 추추출물은 대장암 세포주 HCT116과 SW480세포의 생육을 억제 하였으나 열매추출물은 억제활성이 나타나지 않았다. 잎과 가지 추출물은 cell migration과 wound healing assay를 통해 비정상적인 세포증식 억제를 확인하였으며, β-catenin과 TCF4 의 단백질 수준을 감소시켜 비정상적인 Wnt 신호전달을 억제를 통해 대장암세포의 생육을 억제하는 것으로 판단된다. 따라서 댕댕이나무 잎과 가지는 항암을 위한 대체보완소재 및 천연 항암제 개발을 위한 소재로 활용이 가능할 것으로 판단된다.

이상의 연구 결과로 미루어 볼 때, 도깨비부채 잎(RPL)은 β -catenin의 분해 유도를 통해 대장암, 유방암, 폐암, 전립선암 및 췌장암 세포의 생육을 억제하는 것으로 나타났다. 본 결과는 도깨비부채 잎의 항암을 위한 대체보완소재 및 천연 항암제 개발을 위한 소재로 활용이 가능할 것으로 판단된다. 그러나 추가적 연구를 통해 도깨비 부채 잎의 항암 활성물질의 분석연구가 필요할 것으로 사료된다.

본 연구에서는 다양한 약리학적 활성을 가지는 것으로 알려진 감초 열수추출물(GRW)의 항암효능을 알아보기 위하여 인체 방광암 T24 세포에서 생존율 및 증식억제에 미치는 영향과 이와 연관된 apoptosis 유발 여부 및 관련 인자들의 발현 변화를 조사하였다. 먼저 GRW의 처리에 따른 증식억제 정도를 조사한 결과, GRW 처리 농도 의존적으로 생존율 및 증식억제 현상이 나타났으며, 핵의 형태 변화, DNA 단편화 및 apoptosis 유발에 관하여 조사한 결과 역시 GRW 처리 농도 의존적으로 증가됨을 확인할 수 있었다. 이는 GRW의 처리에 의한 암세포의 증식억제 및 형태적 변형이 암세포의 apoptosis 유발과 밀접한 관련이 있음을 시사하여 주는 것으로 사료된다. GRW 처리에 의한 apoptosis 유발에 관여하는 유전자의 탐색을 위하여 apoptosis와 연관성을 가지는 Bcl-2 family에 속하는 유전자의 발현을 조사한 결과 GRW 처리 농도 의존적으로 Bax 단백질의 발현증가와 더불어 Bcl-2 및 Bcl-xL 단백질의 발현감소가 관찰되었다(Fig. 3A). 이는 GRW에 의한 T24 세포의 apoptosis 유발에 Bcl-2 family에 속하는 유전자의 발현 조절이 중요한 역할을 하는 것으로 사료된다. 또한 GRW의 처리에 따른 MMP의 소실은 미트콘드리아 막의 교란이 유발되었음을 의미하는 것으로, 이러한 MMP 값의 변동은 Bcl-2 family 단백질의 발현 변화에 의한 것이라 추정된다. 한편 Apoptosis에 중요한 역할을 하는 것으로 알려진 caspase(-3/-8/–9)의 발현과 이들의 활성을 억제하는 IAP family (XIAP, cIAP-1, cIAP-2)의 발현에 GRW이 어떠한 영향을 미치는지를 조사한 결과, caspase-3, -8 및 -9의 활성형 단백질 발현 및 정량적 활성증가를 확인하였으며, IAP family 속한 3가지 단백질 모두 발현이 감소하는 것이 관찰되었다. 이상의 결과에서 GRW은 외인적 및 내인적 경로의 개시에 핵심적인 역할을 하는 caspase-8 및 -9의 활성을 모두 증가시켰으며, 이에 따른 caspase-3의 활성증가에 의하여 apoptosis가 유발되었음을 알 수 있었다. 이러한 두 경로의 동시 활성화에는 미트콘드리아의 기능 소실과 Bcl-2 및 IAP family의 발현 변화가 관여하고 있었으며, 특히 Bid의 발현 감소는 GRW에 의한 내인적 경로를 증폭시키는 효과로 작용했을 것이라 추정된다. 방광암의 치료에 보다 효과적인 생리활성을 갖는 물질을 발굴하고 그와 관련된 분자 및 세포수준에서의 기전을 밝히는 것이 중요하기에 본 연구의 결과는 향후 GRW로 수행될 추가 실험을 위한 기초자료로서 그 가치가 매우 높을 것으로 사료된다.

In this study, we evaluated the effect of branch (STB) and leave (STL) extracts from Sageretia thea on β-catenin level in human colorecal cancer cells, SW480 and lung cancer cells, A549. STB and STL dose-dependently suppressed the growth of SW480 and A549 cells. STB and STL decreased β-catenin level in both protein and mRNA level. MG132 decreased the downregulation of β-catenin protein level induced by STB and STL. However, the inhibition of GSK3β by LiCl or ROS scavenging by NAC did not block the reduction of β-catenin protein by STB and STL. Our results suggested that STB and STL may downregulate β-catenin protein level independent on GSK3β and ROS. Based on these findings, STB and STL may be a potential candidate for the development of chemopreventive or therapeutic agents for human colorectal cancer and lung cancer.

In this study, we evaluated the anti-cancer activity and potential molecular mechanism of 70% ethanol extracts of branches from Taxillus yadoriki parasitic to Neolitsea sericea (TN-NS-B) against human lung cancer cells, A549. TY-NS-B dose-dependently suppressed the growth of A549 cells. TY-NS-B decreased β-catenin protein level, but not mRNA level in A549 cells. The downregulation of β-catenin protein level by TY-NS-B was attenuated in the presence of MG132. Although TY-NS-B phosphorylated β-catenin protein, the inhibition of GSK3β by LiCl did not blocked the reduction of β-catenin by TY-NS-B. In addition, TY-NS-B decreased β-catenin protein in A549 cells transfected with Flag-tagged wild type β- catenin or Flag-tagged S33/S37/T41 mutant β-catenin construct. Our results suggested that TN-NS-B may downregulate β- catenin protein level independent on GSK3β-induced β-catenin phosphorylation. Based on these findings, TY-NS-B may be a potential candidate for the development of chemopreventive or therapeutic agents for human lung cancer.

Background : As a part of ongoing research to elucidate and characterize anti-cancer nutraceuticals, extracts from many kinds of medicinal plants were tested for their ability to cytotoxicity on cancer cells so far. Datura stramonium is one of the plants known to contain various alkaloids such as hyoscyamine, scopolamine, atropine etc. Traditionally, it has been used as an analgesic, antispasmodic, and central nervous stimulant. Leaves are also known to be effective against asthma, cough, and chronic bronchitis. In this study, cytotoxicity of extracts from D. stramonium on human cancer cell lines A549 (lung), and HepG2 (liver) were evaluated and compared. Methods and Results : The extract was diluted with DMSO in the form of 10, 50, 250, 1,000 μg/ml for final concentration series respectively. The cell viabilities were examined by MTT assay. On HepG2 cell line, extracts of D. stramonium showed dramatic dose-dependent cytotoxicity on 10, 50, 250, 1,000 ㎍/㎖ concentrations series as 88.16%, 78.55%, 55.57%, 23.06% cell viability respectively. On A549 cell line, likewise, same concentration series showed a dose-dependent cytotoxic effect as 96.49%, 96.12%, 68.54%, 20.26% cell viability respectively. On A549 cell line, there was no difference in effect between 10 ㎍/㎖ and 50 ㎍/㎖. Conclusion : Above results, high concentrations of extracts are effective on two cancer cell lines. These results are expected to be used on further studies about the anticancer activity of D. stramonium as basic data. In order to confirm the anticancer effect of D. stramonium, it is anticipated that additional tests will be required to confirm the apoptosis assay and related protein expression.

Background : Turmeric (Curcuma longa L.) and black pepper (Piper nigrum L.) have traditionally been used as Asian medicinal and culinary herb. Curcumin, a major compound of turmeric, has been known to have antitumor activity. However, curcumin is bioavailable because it is rapidly metabolized and released from the body. Therefore, the addition of adjuvants such as piperine, a potent inhibitor of drug metabolism, is one of the ways to increase the bioavailability of curcumin. Methods and Results : The yields of turmeric and black pepper ethanolic extracts (TM and BP) are 18.2 and 8.2% (w/w), respectively. The EC50 values of A549 and NCI-H292 cells exposed to TM were 77.8 ㎍/㎖ and 92.0 ㎍/㎖, respectively. No significant cytotoxicity was observed up to the 400 ㎍/㎖ in the A549 and NCI-H292 exposed to BP. Based on the central composite design, the co-treatment of TM and BP enhanced the cytotoxicity of A549 and NCI-H292 cells. The optimal combination concentration (optimal EC50 value) of TM and BP calculated by the response surface methodology assay were 48.5 and 241.7 ㎍/㎖. The conbination index assay confirmed that the cytotoxic effect at optimal combinatino concentration was due to the synergistic effect. Conclusion : We hypothesized that co-treatment of TM and BP enhanced cytotoxicity more than single treatment of TM against lung cancer cells, and cell death at this time may synergetic cytotoxicity effects associated with curcumin metabolism.

Dendropanax morbifera Léveille (Araliaceae) is an endemic species growing in the south-western part of South Korea and has been used in folk medicine and health functional food. Several studies have indicated that extract of D. morbifera (DP) has cytotoxic activities on a number of human cancers, such as, breast cancer, lung cancer, hepatoma, and chorioepithelioma. Recently, polyacetylene and triterpene compounds have been isolated from the DP and showed to have anti-complement activity. β-Amyrin, α-amyrin, dendropanoxide, and β-sitosterol are isolated from DP. However, its biological activities in cancer have not yet been clearly elucidated. In this study, we evaluated the anti-cancer activity of isolated triterpenoids from the DP leaves by measuring the levels of cytotoxicity against MCF-7 human breast cancer and A549 human lung cancer cells. Six triterpenoids were isolated from the n-hexane fraction of DP leaves along with the known compounds. β-amyrin (1), α-amyrin (2), olean-12-en-3,24 β-diol (3), dendropanoxide (4), β-sitosterol (5), and stigmasterol (6). Compound 3 and 6 were isolated from DP for the first time. Cytotoxic activities of six compounds were evaluated against two human cancer cell lines by using the MTT in vitro assay. Among them, five compounds (1, 2, 4, 5, and 6) showed moderate cytotoxic activities toward the tested cell lines, and were safe to normal cells. Western blot analysis showed a decreased expression of anti-apoptotic protein Bcl-2 and increased levels of pro-apoptotic protein Bax in MCF-7 and A549 cells treated by β-amyrin and α-amyrin. Flow cytometry analysis confirmed that five compounds (1, 2, 4, 5, and 6) treatment increased populations of sub-G1 (apoptosis) phase. The results of the present study revealed that triterpenoids from DP have the potential for further development as anticancer agents.

Lung cancer, the most common malignant disease worldwide, is the predominant cause of cancer deaths, particularly amongst men. Therefore, various researchers have focused on the growth inhibitory effects of medicinal plants used in traditional Korean medicine. This study aimed to investigate the growth inhibitory effects of ethanol extracts of Rubiae radix, Inulae flos, Nelumbinis receptaculum, Astilbe radix, and Lagerstroemia flos on NCI-H1229 cells. Method and Results: The viability of NCI-H1229 cells was evaluated in vitro using an MTS assay. Treatment with the ethanol extracts of the selected medicinal plants at 500 ㎍/㎖ reduced NCI-H1229 cell viability and increased apoptotic cell death and caspase-3 activation. In addition, treatment with ethanol extracts of Inulae flos and Astilbe radix increases DNA fragmentation, as measured by the TUNEL assay. Conclusions: These results indicated that ethanol extracts of Rubiae radix, Inulae flos, Nelumbinis receptaculum, Astilbe radix, and Lagerstroemia flos exhibited growth inhibitory effects, inducing apoptotic cell death, DNA fragmentation and caspase-3 activation in NCI-H1229 cells. Therefore, these medicinal plant extracts may be used in the development of natural medicines to inhibit the growth of lung cancers. However, further study is needed to determine the active ingredients of the ethanol extracts from medicinal plants that are reposible for the inhibitory effect on lung cancer cell grwoth.

In this study, we investigated the effect of the extracts from Vaccinium oldhamii on cell proliferation and the regulatory mechanisms of cyclin D1 protein level in human cancer cells. The branch extracts from Vaccinium oldhamii (VOB) showed higher inhibitor effect against the cell growth than leave extracts (VOL) and fruit extracts (VOF) in human colorectal cancer, breast cancer, prostate cancer, non-small lung cancer, pancreatic cancer and liver cancer cells. In addition, VOB decreased cyclin D1 level at both protein and mRNA level. MG132 treatment attenuated VOB-mediated cyclin D1 downregulation. A point mutation of threonine-286 to alanine attenuated cyclin D1 degradation by VOB. In addition, the inhibition of nuclear export by leptomycin B (LMB) attenuated cyclin D1 degradation by VOB. But, the treatment of PD98059 (ERK1/2 inhibitor), SB203580 (p38 inhibitor), SP600125 (JNK inhibitor), LiCl (GSK3β inhibitor), LY294002 (PI3K inhibitor) or BAY 11-7082 (IκK inhibitor) did not affect VOB-induced cyclin D1 degradation. In conclusion, VOB induced cyclin D1 degradation through redistribution of cyclin D1 from the nucleus to cytoplasm via T286 phosphorylation of cyclin D1, which resulted in the inhibition of cancer cell proliferation.

Patients with type II diabetes mellitus are more susceptible to colorectal cancer (CRC) incidence than non-diabetics. The anti-diabetic drug metformin is most commonly prescribed for the treatment of this disease and has recently shown antitumor effect in preclinical studies. The aberrant mutational activation in the components of RAS/RAF/MEK/ERK and PI3K/AKT/mTOR signaling pathway is very frequently observed in CRC. We previously reported that metformin inhibits the phosphorylation of ERK and BEZ235, a dual inhibitor of PI3K and mTOR, has anti-tumor activity against HCT15 CRC cells harboring mutations of KRAS and PIK3CA. Therefore, we hypothesized that simultaneous inhibition of two pathways by combining metformin with BEZ235 could be more effective in the suppression of proliferation than single agent treatment in HCT15 CRC cells. Here, we investigated the combinatory effect of metformin and BEZ235 on the cell survival in HCT15 CRC cells. Our study shows that both of the two signaling pathways can be blocked by this combinational strategy: metformin suppressed both pathways by inhibiting the phosphorylation of ERK, 4E-BP1 and S6, and BEZ235 suppressed PI3K/AKT/ mTOR pathway by reducing the phosphorylation of 4E-BP1 and S6. This combination treatment synergistically reduced cell viability. The combination index (CI) values ranged from 0.44 to 0.88, indicating synergism for the combination. These results offer a preclinical rationale for the potential therapeutic option for the treatment of CRC.

Background : Although the inhibitory effect of mistletoe on cancer cell growth has been reported, the underlying mechanisms to explain its anti-proliferative activity are not fully studied. Thus, we elucidated the potential molecular mechanism of the branch from taxillus yadoriki (TY) parasitic to Neolitsea sericea (NS) (TY-NS-B) for the anti-proliferative effect. Methods and Results : In comparison of anti-proliferative effect of TY from the host trees such as Cryptomeria japonica (CJ), Neolitsea sericea (NS), Prunus serrulata (PS), Cinnamomum camphora (CC) and Quercus acutissima (QA), TY-NS showed higher anti-cell proliferative effect than TY-CJ, TY-PS, TY-CC or TY-QA. In addition, the anti-proliferative effect of branch from TY from all host trees was better than leaves. Thus, we selected the branch from Taxillus yadoriki parasitic to Neolitsea sericea (TY-NS-B) for the further study. TY-NS-B inhibited the cell proliferation in the various cancer cells and downregulated cyclin D1 protein level. MG132 treatment attenuated cyclin D1 downregulation of cyclin D1 protein level by TY-NS-B. In addition, TY-NS-B increased threonine-286 (T286) phosphorylation of cyclin D1, and the mutation of T286 to alanine (T286A) blocked cyclin D1 proteasomal degradation by TY-NS-B. But the upstream factors related to cyclin D1 degradation such as ERK1/2, p38, JNK, GSK3β, PI3K, IκK or ROS did not affect cyclin D1 degradation by TY-NS-B. However, LMB treatment was observed to inhibit cyclin D1 degradation by TY-NS-B, and T286A blocked cyclin D1 degradation through suppressing cyclin D1 redistribution from nucleus to cytoplasm by TY-NS-B. In addition, TY-NS-B activated CRM1 expression. Conclusion : Our results suggest that TY-NS-B may suppress cell proliferation by downregulating cyclin D1 protein level through proteasomal degradation via T286 phosphorylation-dependent cyclin D1 nuclear export. These findings will provide the evidence that TY-NS-B has potential to be a candidate for the development of chemoprevention or therapeutic agents for human cancer.