We have previously shown that 5’-nitro-indirubinoxime (5’-NIO) has potent anti-tumor effect in various human cancer cells. But the potential anti-invasive effect of 5’-NIO in salivary gland cancer has not been studied yet. The goal of this study is to evaluate the effect of 5'-NIO on salivary gland adenocarcinoma SGT cell adhesion and migration to Type I collagen treatment. Western blot, adhesion and migration assay were performed to evaluate the impacts of 5’-NIO on the expression of MMP-2/-9 and its upstream signaling molecules after treatment of type І collagen. SGT cell adhesion to type I collagen is significantly suppressed by 5’-NIO. 5’-NIO decreased expression of β1 integrin, phosphorylation of FAK, MMP-2/ -9 compared with type I collagen treatment. In addition, 5’-NIO inhibited the migration of SGT cells treated with type I collagen. These results suggest that 5’-NIO could effectively inhibit the invasion and migration of human SGT cells by downregulating the expression of β1 integrin and MMP-2/-9 and phosphorylation of FAK, Akt, and Erk. Adhesion and migration to type I collagen of SGT cells can be influenced through 5’-NIO..

Cyclosporine A (Cs A) which is a highly lipophilic cyclic undecapeptide mainly used for its immunosuppressive properties exert a wide spectrum of biological activities including fungicidal antiproliferactive, anti-inflammatory and chemotherapuetic effects. Human salivary gland adenocarcinoma is very aggressive characteristics, which is need to get the effective chemotherapuetic methods. Subconfluent SGT cell cultures have been treated with CsA at in vivo relevant concentrations for 24h. MTT assay for cellular proliferation of cultured SGT cell line has been performed and TGase 1 activity assay for cellular differentiation has been detected in the CsA-treated samples. It suggested that CsA could have an inhibitory effect in the proliferation of SGT cell line but no in the differentiation.

The human embryonic-lethal abnormal vision-like protein, HuR, stabilizes mRNA containing adenine- and uridine- rich elements in their 3’untranslated region. Because cyclooxygenase-2 (COX-2) mRNA is a cellular transcript that contains an adenine- and uridine-rich element, it can be regulated by the HuR protein. In this study, we examined the relationship between COX-2, HuR, MVD, and the clinicopathological parameters. Nineteen out of 43 cases of HNSCC showed high level of COX-2expression, and 68% of these patients showed high COX-2 immuno-reactivity indicating the strong expression of the cytoplasmic HuR protein. Also, MVD expression in the cases with high COX-2 expression was higher than in the cases with low COX-2 expression. These results suggest a strong correlation between the overexpression of cytoplasmic HuR and COX-2 expression in HNSCC, and that COX-2 is associated with MVD in HNSCC. In conclusion, COX-2 regulated by cytoplasmic HuR may be a good tumor angiogenic factor in HNSCC.

본 연구에서는 참외 추출물의 항암활성에 대해 알아보기 위해 참외를 부위별로 나누어 quinone reductase 유도활성과 다양한 간암세포에서의 증식 억제활성을 조사하였다. 참외 꼭지와 참외 줄기 잎 부위에서 농도의존적으로 QR 유도활성이 증가하였고, 200μg/mL 농도에서는 각각 3.9, 1.5배의 유도활성을 나타내었다. 암세포 사멸 활성 측정법을 통한 항암활성 평가 실험에서 마우스 유래의 간암세포인 Hepa1c1c7 세포에 대해서 조사한 결과 꼭지와 줄기 잎 부위에서 높은 암세포 독성을 보였다. 이러한 결과를 기초로 인체유래의 암세포에 대한 항암활성을 평가하기 위해 인체유래 간암 세포주인 HepG2에 대한 세포 증식 억제활성을 농도별로 조사하였다. 꼭지와 줄기 잎 부위 모두 인체유래 간암 세포에 대해 증식 억제효과를 보여주었지만, 특히 꼭지 부위는 최고농도에서 60.3%의 높은 증식 억제효과를 보였다. 그러나 마우스 유래의 간암세포에 대한 활성보다 인체유래 간암세포에 대한 활성이 낮게 나타났다. 참외의 꼭지 추출물에서 QR 유도활성과 항암활성을 확인함으로써 향후 참외 비가식 부위의 기능성 소재로의 이용화에 대한 연구가 필요할 것으로 생각된다.

본 연구에서는 효소의 활성을 증진시킬 수 있는 새로운 방법의 개발을 위하여 정제된 polygalacturonase(PGase)에 guar gum, xanthan gum, locust bean gum 등과 같은 교질물질을 첨가함으로써 효소의 활성을 특이적으로 향상시킬 수 있는 방법 조사하였다. 정제된 PGase를 0.2%의 상술한 교질물질을 함유하는 동일한 완충용액에 잘 혼합시켜 30oC에서 배양하여 상대활성을 구한 결과, guar gum 첨가 시 는 PGase의 상대활성이 105%가 증가 되었으며, xanthan gum 첨가 시는 87%, locust bean gum 첨가 시는 90%가 증가되어 상술한 교질물질들이 효소의 활성 촉진제로서의 기능이 있음을 확인할 수 있었다. Guar gum 처리에 의해 당근 단세포 생성 반응을 수행한 결과 모든 반응시간에서 guar gum을 가했을 때가 PGase만을 가하여 단세포화 반응을 수행한 경우 보다 높은 수율을 보였으며, 단세포화 반응 2시간 경과 후에 대조군 대비 13%의 가장 높은 수율 향상을 보였다. 이상의 결과에서 PGase에 guar gum, xanthan gum, locust bean gum 등과 같은 교질물질을 첨가함으로써 효소의 활성을 특이적으로 향상시킬 수 있으며, 이를 이용한 단세포 당근의 제조에 있어서 효소반응의 수율 증진을 위한 방법으로 활용될 수 있음을 확인하였다.

Numerous bone cell culture models have been presented by the development of isolation and culturing techniques of cells. The culture of osteoblast-like cells of human origin with a specific osteoblastic phenotype has become an important experimental model in bone biology. Recently, it has become increasingly popular to utilize bone marrow cultures because these cultures are therefore thought to represent earlier stages of the osteoblast differentiation pathway. There is no report about culturing normal human osteoblast from oral and maxillofacial area. Primary cultured cells from oral and maxillofacial cancellous bone were analyzed by morphologic features, total DNA contents, ALP, osteocalcin and von Kossa staining positivity. The purpose of this study were to culture the cell population from oral and maxillofacial cancellous bone and to analyze the phenotypic expression of cultured normal human osteoblast by the bone marrow isolation technique. Growth curve of NHost showed about 45hrs of doubling time and about 70μ g/well of total DNA content. NHost showed spindle shaped cytoplasm with ovoid nucleus under preconfluency and after cellular differentiation, they formed irregular numerous nodules from stratified cellular layers under D medium. ALP activity was about 2 folds higher under control medium with 10nM 1,25(OH)2D3 than that under control medium. Osteocalcin expression was about seven folds higher under control medium with 100nM 1,25(OH)2D3 than that under control medium. Scattered mineralized nodules stained by von Kossa method were seen on the cellular layer under D medium. It suggested that NHost might be established from oral and maxillofacial area by characteristic cellular shape, ALP activity, osteocalcin expression and numerous mineralized nodules.

Epigallocatechin-3-gallate (EGCG) and theaflavins (TF) are polyphenols included in green and black teas, respectively. Both green and black teas have been studied for their potential health benefits for cancer. Hypoxia-inducible factor (HIF) has been implicated multiple physiological and pathophysiological pathways, particularly, oncogenesis. But, the molecular pathways that govern the cell response to EGCG are not fully elucidated. The present study investigated the intracellular mechanism in oral squamous cell carcinoma (OSCC) cells treated with EGCG, focusing on HIF-1 expression and its effect on epithelial phenotype. EGCG decreased phosphorylated Raf-1 protein in YD 8 OSCC cell, but B-raf protein was not affected at all by EGCG and TF. In addition, we here found that EGCG regulated HIF-1α expression independent of Raf-1 protein. Taken together with our previous result, the result imply that EGCG is attributed to the HIF-1α expression via Raf/MEK/ERK pathway, and the HIF-1α expression is associated with the change of epithelial phenotype in OSCC cell.

Melanoma is the most serious type of skin cancer as a malignant tumor of melanocytes. In this work, the syntheses of a novel series of benzaminoquinoline derivatives 1a-c and their antiproliferative activities against A375 human melanoma cell line were described. All the compounds (IC50=0.78-1.02μM) showed superior antiproliferative activities to Sorafenib (IC50=5.58μM) as a reference compound. These results suggested that benzaminoquinoline derivatives have potentials as a therapeutic agent for the treatment for melanoma.

The trace element nutrient selenium discharges its well-known nutritional anti-tumor activity. Converging data from epidemiological, ecological and clinical studies have shown that selenium can decrease the risk for some types of human cancers, especially those of the prostate, lung, and colon. Mechanistic studies have indicated that selenium has many desirable attributes of chemoprevention targeting cancer cells through DNA single strand breaks, the induction of reactive oxygen species. However, there is no reports about the relationship between methylseleninic acid (MSeA), one of methylselenol metabolites and cell cycle arrest in LNCaP human prostate cancer cells. Our data showed that MSeA arrested G1/S pahse of cell cycle arrest and inhibited DNA synthesis in LNCaP cells and those cellular events by MSeA were due to the induction of p27 protein which is a well-known cyclin-dependent kinase inhibitor. Taken together, cell cycle arrest occurred by MSeA may contribute to the growth-inhibition of prostate cancer cells.

Two-pore domain 칼륨() 통로는 흥분세포 및 비흥분세포의 안정막 전압을 일정하게 유지하는데 관여한다. 그러나 생식세포 및 생식기관에서 발현되는 통로의 분포영역 및 그 기능에 대해서는 연구자들에 의해 아직 정리되지 못하였다. 본 종설에서는 통로의 생식세포 및 생식기관에서 발현, 분포 및 생리학적 의의를 논하였다. 통로는 인간 영양막세포, 자궁근층, 태반혈관계, 자궁평활근조직, 태반융모조직 및 임신자궁조직에서 발현되어 임신에 있어서 관련성을 제

In this study, we aimed to determine whether the evaluated markers of cell death could be found at particular developmental stages of normal porcine in vitro fertilization (IVF) embryos. We investigated the characteristics of spontaneous and induced apoptosis during preimplantation development stages of porcine IVF embryos. In experiment 1, to induce apoptosis of porcine IVF embryos, porcine IVF embryos at 22h post insemination were treated at different concentration of actinomycin D (0, 5, 50 and 500 ng/ml in NCSU medium). Treated embryos were incubated at in 5% , 5% for 8h, and then washed to NCSU medium and incubated until blastocyst (BL) stage. We examined cleavage rate at 2days and BL development rate at 7days after in vitro culture. A significantly lower rate of cleavage was found in the 500 ng/ml group compared to others (500 ng/ml vs. 0, 5, 50 ng/ml; 27.8 % vs. 50.0%, 41.2%, 35.9%), and BL formation rate in 500 ng/ml was lower than that of others (500 ng/ml vs. 0, 5, 50 ng/ml; 8.0% vs. 12.6%, 11.2%, 12.6%). In experiment 2, to evaluate apoptotic cells, we conducted TUNEL assay based on morphological assessment of nuclei and on detection of specific DNA degradation under fluorescence microscope. This result showed that apoptosis is a normal event during preimplantation development in control group (0 ng/ml actinomycin D). A high number of BL derived control group contained at least one apoptotic cell. Actinomycin D treated BLs responded to the presence of apoptotic inductor by significant decrease in the average number of blastomeres and increase in the incidence of apoptotic cell death. In 500 ng/ml group, the incidence of apoptosis increased at 4-cell stage and later. This result suggested that apoptosis is a process of normal embryonic development and actinomycin D is useful tool for the apoptosis study of porcine preimplantation embryos.

Changing Proteins in Granulosa Cells during Follicular Development in Pig In-Soon Chae1, Dong-Min Jang3, Hee-Tae Cheong2, Boo-Keun Yang1 and Choon-Keun Park1,† 1College of Animal Life Sciences, 2School of Veterinary Medicine, Kangwon National University, Chuncheon 200-701, Korea 3CHA Stem Cell Institute, CHA Hospital, Seoul 463-712, Korea ABSTRACT This study analyzed change of proteins in granulosa cells during the porcine follicuar development by proteomics techniques. Granulosa cells of the follicles, of which the diameter is 2～4 mm and 6～10 mm, were collected from ovary of slaughtered pig that each follicle of diameter 1～4 mm and 6～10 mm. We extracted glanulosa cell proteins by M-PER Mammalian Protein Extraction Reagent. Proteins were refined by clean-up kit and quantified by Bradford method until total protein was 200 μl. Immobilized pH gradient(IPG) strip used 18 cm, 3～10 NL. SDS-PAGE used 10% acrylamide gel. After silver staining, Melanie 7 and naked eye test were used for spot analyzation. Increasing proteins in glanulosa cell of 6～10 mm follicle were 7 spots. This spots were analyzed by MALDI-TOF MS and searched on NCBInr. In results, 7 spots were similar to zinc/ling finger protein 3 precursor (RING finger protein 203), angiomotin, heat shock 60 kDa protein 1 (chaperonin) isoform 1 (HSP60), similar to transducin-like enhancer protein 1 (TLE 1), SH3 and PX domains 2A (SH3PXD2A). Those proteins were related with transfer between cells. Increase of proteins has an effect on follicular development.

This study was conducted to investigate the variation of growth characteristics and reproductive physiology in cloned Hanwoo male calves during growing stage. The hematological parameters, body weight, and plasma hormonal levels, birth to 12 months, were analyzed in the cloned calves (n=3). Differences among treatment means were determined by a student t-test. A probability of P<0.05 was considered statistically significant. The hematological parameters, such as white blood cell, red blood cell, and platelet, were not different in both normal and cloned calves. The difference of body weight, however, was significantly higher in the cloned calves, months (p<0.05) and months (p<0.01), than that of the comparators, respectively. The plasma IGF-1 level was statistically significant in the cloned calves, months, compared to that of the normal calves (p<0.05). However, the plasma testosterone level was not different in both normal and clone calves according to growing stage. Taken together, the cloned Hanwoo male calves are growing faster and maintaining a normal reproductive physiology.

Spermatogonial stem cells(SSCs) only are responsible for the generation of progeny and for the transmission of genetic information to the next generation in male. Other in vitro studies have cultured SSCs for proliferation, differentiation, and genetic modification in mouse and rat. Currently, information regarding in vitro culture of porcine Germline Stem Cell(GSC) such as gonocyte or SSC is limited and is in need of further studies. Therefore, in this study, we report development of a successful culture system for gonocytes of neonatal porcine testes. Testis cells were extracted from 10～14-day-old pigs. These cells were harvested using enzymatic digestion, and the harvested cells were purified with combination of percoll, laminin, and gelatin selection techniques. The most effective culture system of porcine gonocytes was established through trial experiments which made a comparison between different feeder cells, medium, serum concentrations, temperatures, and O2 tensions. Taken together, the optimal condition was established using C166 or Mouse Embryonic Fibroblast(MEF) feeder cell, Rat Serum Free Medium(RSFM), 0% serum concentration, 37℃ temperature, and O2 20% tension. Although we discovered the optimal culture condition for proliferation of porcine gonocytes, the gonocyte colonies ceased to expand after one month. These results suggest inadequate acquirement of ingredients essential for long term culture of porcine GSCs. Consequently, further study should be conducted to establish a successful long-term culture system for porcine GSCs by introducing various growth factors or nutrients.