Cordyceps militaris mycelium extracts containing high amounts of cordycepin were evaluated in vitro for their antiinflammatory and tumor cell growth-inhibitory activities. All extracts dose dependently inhibited the increased production of inflammatory mediators including reactive oxygen species (ROS), nitric oxide (NO), and β-hexosaminidase in lipopolysaccharide (LPS)-stimulated inflammatory cells. All extracts were evaluated for anti-proliferative activity against normal RBL-2H3 cells and diverse types of cancer cell lines, including HCT, MC5-7, U-87MG, AGS, and A549 cells. The extract showed the strongest growth inhibition (IC50 = 28.13 μg/mL) relative to vehicle-treated control cells against fibrosarcoma (MC5-7). We have demonstrated anti-inflammatory activity of C. militaris via inhibition of NO, ROS production, and β-hexosaminidase release in activated cells. C. militaris mycelium extract was also evaluated mechanistically and found to exert six types of anti-cancer activity, confirming its pharmacological potential. Our study suggests C. militaris use as a potential source of anti-inflammatory and anticancer agents. C. militaris may also be considered a functional food.

Fucoidan is a sulfated polysaccharide that is purified from brown algae, such as Fucus vesiculosus. This compound has multiple biological activities including immune-stimulating, and anti-viral activities. We recently demonstrated that the cytotoxicity of fucoidan can be dependent on the batch of its production and its molecular weight. In a previous study, fucoidan B exerted cytotoxicity toward mouse spleen cells. To confirm the biological activity of Fucoidan B, we cultured HL-60 cells, a human leukemia cell line, and treated then with fucoidan. The metabolic activity of the HL-60 cells decreased in response to treatment by fucoidan. Moreover, the morphology of HL-60 treated by fucoidan changed. To investigate the fucoidan’s effects, we analyzed the size and level of Annexin V/propidium iodide staining of HL-60 cells using a flow cytometer. Fucoidan consistently induced cell death, including apoptosis of HL-60 cells. As potential mechanisms, fucoidan destabilized the mitochondrial membrane potential and altered the production of reactive oxygen species in HL-60 cells. Taken together, these results suggest that fucoidan has anti-tumor activity on HL-60 cells via destabilization of mitochondrial membrane potential. The present study demonstrates that fucoidan can be used as an anti-cancer agent for leukemia.

The lesser paper wasp, Parapolybia varia, belongs to large subfamily Polistinae and is distributed in Middle East, the Indo-Papuan region and East Asia. P. varia is known to become aggressive when disturbed for defending their colonies, resulting in fatal envenomation. Vespid chemotactic peptide (VCP) and vespakinin have recently been determined to be the top two genes most abundantly transcribed in venom glands of P. varia. To investigate the pharmacological and toxicological properties of VCP and vespakinin, their antitumor, antimicrobial, and cytotoxic activities were evaluated. VCP exhibited a significantly high antitumor activity against ovarian tumor cell SK-OV-3 at 100 M. VCP also showed higher hemolytic activity than vespakinin. Antimicrobial activity was only observed with VCP against yeast Candida albicans at 1 mM. Since VCP showed a relatively low hemolytic activity but a considerable level of antitumor activity, it has further merits to be exploited as a potential antitumor agent with reduced side effects on normal cells.

Codonopsis lanceolata L. (Campanulaceae) has long been used in traditional Korean medicine to treat bronchitis, cough, and inflammatory diseases, however, the efficacy of anti-tumor activities remains to be defined. In this study the effects of Codonopsis lanceolata (C. lanceolata) on proliferation, migration and adhesion in lung (A549, H1299) and ovarian cancer (SKOV-3) cells were investigated. To assess and compare the pharmacological effects and production places of C. lanceolata, the ethanolic extracts of C. lanceolata from different places in Korea (Hongseong, Yecheon, Yeongwol, Yanggu, Gangjin, and Hoengseong) were prepared. The extract from Hoengseong county did have only marginal anti-proliferative activity in all the cell lines tested, however, other extracts had little or no effect on cell proliferation. The extracts from Hongseong, Gangjin or Hoengseong county had partial anti-migratory activity in lung cancer cells, but not in ovarian cancer cells. In addition, the extract from Hoengseong county had partial anti-adhesive activity in ovarian cancer cells, however, other extracts did not affect cell adhesion in both lung and ovarian cancer cells. Taken together, these findings provide the first description of anti-tumor efficacy of C. lanceolata from different production places in Korea, and suggest that C. lanceolata from Hoengseong county may have therapeutic potential in lung and ovarian cancers.

1,1- Bis(3’-indolyl)-l-(p-methoxyphenyl)methane (DIM- C- pPhOCH.3) is a methylenc - substituted diindolylmethanes (C-DIM) ana log that acti vates the orphan receptOl‘ nerve growth factor-induced-B (NGFI-B, Nur77) , RNA inteference studies with small inhibitory RNA for Nur77 demonstrate that DIM-C-pPhOCH:J induces Nur77-dependent and - independent apoptosis, and this study has focused on delineating the Nur77-independent proapoptotic pathways induced by the C-DIM analog DIM-C-pPhOCH3 induced caspase-dependent apoptosis in RKO colon cancer cells through decreased mitochondrial membrane potential which is accompanied by increased mitochondrial bax/bcl-2 ratios and release of cytochrome c into the cytosol DlM-C一pPhOCH.3 also induced phosphatidylinositol-3-kinase-dependent activation of early growth response gene-l whi ch, in turn, induced expression of the proapoptotic nonsteroidal anti-inflammatory drug- activated gene-l (NAG- l) in colon tumors in athyrnic nude mice bearing RKO cells as xenografts, DIM-C-pPhOCH.3 also activated the extrinsic apoptosis pathway through increased phosphorylation of c- jun N-terminal kinase which, in turn, activated C/EBP homologous transcription factor (CHOP) and death receptor 5 (DR5) , Thus, the effectiveness of DIM-C-pPhOCH.3 as a tumor growth inhibitor is through activation of Nur77-dependent and -independent pathways

Indirubin is the ac ti ve ingredi ent of a traditional Chinese herbal medicine, Danggui Longhui Wan, used for t he t reatment of chronic myelocytic leukemia Here, we report that novel indirubin ,- 11‘ ivative‘ 5’ - nitro-indirubinoxime (5’ NIO) , has potent anti- proliferative activity on va rious human cancel‘ cells and oncogenic RK3E- ras rat kidney cells with ha lf- inhibi tory concentrati ons (1C50) ra nging from 1- 12 M, Treatment with indirubin derivative induced the activation 01' caspase 7 rollowed by apoptos is in RK3E- ras cells. lndirubin derivative showed strong anti-tumor activity in rat solid and oral tUll10r models , Direct inj ection of indirubin deri vative every other day for 10 days induced signifi cant inhi bition of tumor growth in Sprague-Dawley rats bearing RK3E- ras-induced tumors Histologically. t reatment with indiru bin de ri vative caused s ignifi cant inhi bit ion of tumor formation with increased apoptosis and decreased tumor cell prolife ration, These f indings provide the potent ial va lue of indirubin deri vative a s a novel candidate for ant i-tumor agents

lndil‘ ubin is the active ingredient of Danggui Longhui Wan‘ a mixture of herbal medici l1e t hat is used to treat chronic myelocytic leukemia in tradi t ional Chinese medicine‘ He re, we show that new indirubin deri vatives 5' -ni tro-indirubinoxime, 5' -f1 uoro-indiru binoxime and 5’ -tri rnethylacetamino-i nd i 1'1.1 bi noxi me‘ have potent a n ti- proliferative activity on various human cancer cells and oncogenic RK3E-ras rat kidney cells with lC50 con centration ranging from 1-25 μ M, When the RK3E-ras cells were treated with indirubin derivatives for 24 h, the activity of caspase-3 and caspasc-7 was induccd followed by apoptosis , 011 the other hand, the activity of SAPK/JNK was inhi bited over the same period , lndirubin deri vatives a lso s howed strong ant i-tumor activity in rat s이 id and oral tumor models The inhibition of tlUl10r g:rowth was observed in animals beaJ‘ing RK3E-ras-induced turnor given subcutaneous dose of 100 mg/kg every other day for 10 days, Histologically, μeatment of indirubin derivatives caused significant inhibition of tumor formation associated wi th increased apoptosis and decreased proliferation of tumor cell s , These findings provide the potential value of indirubin derivatives as a novel candi date fOl 없l tJ -cancer agents ,

Endocrine disruptors are exogenous chemicals that their endocrine disrupting effects mediated by androgenic signaling plays crucial roles in the control of development and several androgen-related diseases. However, there are no authorized in vitro screening and testing methods to evaluation of (anti-)androgenic activity. To find out a better in vitro cell line model, we have previously reported that 22Rv1 cells, a human prostate cancer cells contained functional Androgen Receptor (AR), might be an appropriate model for the evaluation of (anti-)androgenic endocrine disruptors. Based on this result, we developed a stable 22Rv1/mouse mammary tumor virus (MMTV) cell line to test AR-mediated transcriptional activation (TA). Using 22Rv1/MMTV cells, we established the test protocol and optimized the testing condition for AR-TA assay. In this study, we performed the inter-validation assay by four different laboratories to evaluate the 20 coded chemicals which were selected from the ICCVAM list (ICCVAM, 2003) or academic articles that exhibited exact (anti-) androgenic activity. The statistical analysis of the results of the inter-laboratory validation study revealed that there was reproducibility between the four participating laboratories. In conclusion, 22Rv1/MMTV AR-TA assay might be a quick and relatively inexpensive method, which can be used to screen large numbers of chemicals for their potential to activate or inhibit AR-mediated gene transcription. Furthermore, it will provide mechanistic data relevant to understanding adverse reactions observed in intact organisms.

본 연구는 길경 사포닌 추출물(PGS)를 이용하여 대식세포의 면역조절능력을 평가하였으며, 탐식작용, 항암작용, 항염증 작용에 모두 유의적인 효과를 나타내었다. 특히, 본 연구에서는 농도의존적으로 매우 유의적이게 나타난 PGS의 항암작용 기전을 측정하기 위하여 암세포 독성 물질로 알려진 NO 분비량을 측정하였으며, PGS에 의해 NO의 생성을 증가함을 확인하였다. 또한, PGS가 NO 생성 억제제 NIL을 함께 처리하였을 때 항암효과가 나타나지 않게 됨을 재확인함으로써, PGS 10 μg/mL에서 나타낸 대식세포의 항암효과는 일부 NO 생성 및 분비에 의한 작용 기전임을 보여주었다. PGS의 면역조절작용 중 항염증효과 실험에서는 PGS가 염증환경에서 과도하게 분비된 NO를 다소 억제하는 경향을 보였으나, 항염증조절에서 대표적인 물질로 알려진 TNF-α 조절에는 효과를 나타내지 않았다. PGS가 염증환경에서의 TNF-α억제조절에는 영향을 미치지 않았으나 TNF-α는 항암물질로도 알려져 있으므로 향후 PGS의 항암효과에 대한 연구에서 TNF-α의 생성에 관한 연구는 NO를 매개하는 항암 효과 외에 다른 기전을 설명해줄 수 있을 것으로 보인다. 이러한 결과들은 PGS가 항암요법의 보조제 및 면역보조제로써의 활용에 개발 가능성이 있다는 것을 보여준다.

To evaluate the anti-tumor activity of Flammulina velutipes extract, we used an in vitro wound-healing assay, and an in vivo approach using a mouse melanoma model. Wound-healing activity in B16 cells was affected by the extract in a dose-dependent manner, indicating that the extract had anti-metastatic activity. The extract also exhibited strong anti-tumor activity against lung cancer when B16 cells were injected into mouse veins together with B16 melanoma cells. The results indicatethat the Flammulina velutipes extract decreased B16 cancer cell growth by inhibition of cell migration both in vitro and in vivo.

To search for immunoactive natural products exerting anti-inflammatory activity, we have evaluated the effects of the ethanol extracts of Rubus coreanus Miq. (ERC) on lipopolysaccharide-induced nitric oxide (NO), tumor necrosis factor-α (TNF-α), and Interferon-γ (IFN-γ) production by RAW 264.7 macrophage cell line. Our data indicate that this extract is a potent inhibitor of NO production and it also significantly decreased IFN-γ and TNF-α production. Consistent with these results, the protein level of inducible Nitric Oxide Synthase (iNOS) and cyclooxygenase-2 (COX-2) was inhibited by ethanol extracts of ERC in a dose-dependent manner. These results suggest that ERC may exert anti-inflammatory and analgesic effects possibly by suppressing the inducible NO synthase and COX-2 expressions.

이전에 보고되었던 복분자, 당귀의 추출물의 항암 및 면역활성에 대한 2차 생리활성 검증을 위한 in vitro 실험으로 Sarcoma-180에 의해 유발된 복수암, 고형암에 대한 항암 효과 및 면역과 관련하여 항스트레스 활성에 대한 실험을 실시하였다. Sarcoma-180에 의해 유발된 복수암과 고형암에 대해 마우스의 체중을 20일간 측정하여 체중의 변화를 관찰한 결과 당귀, 복분자 추출물 모두 대조구에 비해 체중의 변화가 적은것으로 나타났다. 대조구는 20일째 47.3g으로 나타났고, 당귀는 45.8g, 복분자는 42.1g으로 나타나 두 가지 시료 모두 항암 효과가 있는 것으로 나타났고, 이 중 복분자가 더 높은 항암 효과를 나타낸 것으로 나타났다. 면역활성에 대한 2차 검증 실험의 하나인 항스트레스 활성을 측정한 결과 복분자와 당귀 추출물 모두 스트레스에 좋은 효과를 나타내었다. 항스트레스 활성 측정은 혈액내 스트레스와 관련 있는 cholesterol과 glucose를 측정하였고, 면역장기인간, 부신 및 비장의 장기의 무게를 측정한 결과 당귀, 복분자 모두 대조구에 비해 스트레스에 대한 좋은 효과를 나타내었다. Cholesterol과 glucose측정 결과 복분자 투여 시 두 가지 모두 standard control에 가까운 결과를 나타내었다. 장기 무게를 측정한 결과 복분자, 당귀 추출물 모두 대조구에 비해 장기 무게가 감소하는 경향을 나타내었다. 당귀와 복분자는 이전에 여러 가지 좋은 효능을 가지고 있다고 보고되었고, 그중에서 항암 및 면역활성에서도 좋은 효과를 나타낸다는 보고가 있었다. 이에 대한 2차 검증 실험으로 in vitro 실험을 통하여 항암 활성과 면역활성과 관련이 있는 항스트레스 활성을 측정함으로서 기능성 식품 개발에 기초적인 근거를 제공할 수 있는 것으로 생각되어진다.

한의학에서 천마는 여러 치료의 목적으로 사용이 되는 한약재이다. 본 연구는 천마의 항산화활성과 항암 효과를 검증하기 위하여 실시하였다. 천마와 이를 이용한 식음료를 이용하여 항산화활성과 암세포를 이용한 세포의 운동성을 측정하였다. 항산화활성의 경우 DPPH, FRAP, Hydroxyl radical assay를 실시하였으며, 항암효과를 확인하기 위하여 wound / invasion assay를 실시하였다. 실험 결과 천마의 항산화 활성은 매우 뛰