• Asian Pacific Journal of Cancer Prevention
• /
• 제15권6호
• /
• pp.2655-2661
• /
• 2014

Background: Talin-1 is a cytoskeleton protein that participates in cell migration and plays a role in tumor formation, migration, and metastasis in different types of cancer. Chinese investigators have observed that the levels of Talin-1 protein and mRNA expression in HCC tissues are significantly lower than in the adjacent non-cancerous tissue. However, Japanese investigators have reported that Talin-1 is upregulated in HCC. Tln2 as homologous gene of Tln-1, which encodes a very similar protein, but the role of Talin-2 is very little known in primary liver cancer (PLC). We investigated whether the expression of Talin-1 in PLC may be associated with the histological subtype as well as the role of Talin-1 in tumor cell invasion and migration using human hepatocellular carcinoma cell lines. Materials and Methods: We measured the mRNA expression levels of Talin-1 and Talin-2 in five human liver cancer cell lines and normal human liver cell ($LO_2$ cell line) by real-time PCR and the protein expression levels of Talin-1 by Western blot. Migration and invasion of the cells were assessed using transwell assays and cell scratch experiments, respectively, and proliferation was assessed by soft AGAR colony formation. Results: Talin-1 and Talin-2 expression differed significantly between the five human liver cancer cell lines and $LO_2$ cell line (p<0.05). Compared with the $LO_2$ cell line, the invasion and migration capabilities of the five cancer cell lines differed significantly (p<0.05). Similarly, the colony-forming ability differed (p<0.05). Conclusions: High levels of Talin-1 expression are correlated with reduced invasion and migration as well as decreased malignancy in human liver cancer cell lines; the suppression of Talin-1 promotes invasion and migration. In addition, Talin-2 may be correlated with invasion and migration in human hepatocellular carcinoma.

• Journal of Periodontal and Implant Science
• /
• 제25권2호
• /
• pp.210-221
• /
• 1995

The purpose of this study was performed to investigate the mineralization and differentiation of osteobalsts for bone regeneration in vitro and the effect of rate of the composition in periodontal cells on mineralization. For this study, healthy gingival tissues were surgically obtained from the patients during 1st premolar extraction for the purposes of orthodontic treament. Gingival tissue was washed several time with Phosphate buffered saline contained high concentration of antibiotics and antifungal agent, and cultured in Dulbecco's Modified Eagle's Medium(DMEM, Gibco, U.S.A.). Every cell were cultured in state at $37^{\circ}C$, 100% of humidity, 5% of $CO_2$ incubator. Bone marrow stromal cells were isolated from 5-clay-old rat femur with using medium irrigation mathod by syringe. Cell suspension medium were centrifuged at 1500 rpm for 5 min and then cultured in the petri dish. Two kinds of cell were freezed and stocked in the liquid nitrogen tank until experiment. Cell were incubated into the 24 multi-well plate with $5{\times}10^4$cell/well of medium at $37^{\circ}C$, 100% of humidity 5% $CO_2$ incubator for 24 hours. After discarded of the supernatent of medium, O.5ml of medium were reapplied and incubated. And counted the number of cell using the hemocytometer and inverted light microscope. We have measured the number of mineralized nodule with using Alizarin red S. staining in microscope. Furthermore every cell were observed the morphological change between every rate of co-culture of the two kinds of cell. The results were as follows; The rate of proliferation of co-culture cell revealed high rate tendency compared the bone marrow stromal cell only and low growth rate to compared with gingival fibroblast only. The tendency of formation of the mineralized nodule were observed dose-depend pattern of bone marrow stromal cell. It is concluded that the gingival fibroblast may inhibit the formation of mineralized nodule in the culture of the bone marrow stromal cell.

• 환경생물
• /
• 제26권1호
• /
• pp.30-35
• /
• 2008

곰피추출물의 CCD-986sk cell line monolayer (human fibroblast, KCBL-21947)에 대한 피부세포 생리활성효과를 측정하고, 또한 곰피추출물의 Clone M-3 mouse melano-cyte cell line에 대한 melanin formation 저해효과를 측정하기 위해 in vitro레벨에서 실험을 실시하였다. 곰피는 다년생 갈조류의 일종으로 이 종은 한국 연안해역에서 중요한 1차생산자의 역할을 담당하고 있는 종이며 연안 생태계에 있어서 생태학적으로나 수산업상 매우 중요한 위치를 점유하고 있다. 최근 이러한 대형 갈조류에 관한 연구는 주로 생태학적인 관점에서 이루어져 왔으며(Notoya and Aruga 1990), 유주자의 발아와 핵분열(Yabu and Notoya 1985) 등에 관한 연구가 주로 일본의 연구자들에 의하여 보고된 바 있고, 국내에서는 Park et al. (1994)이 부산만 인근해역 곰피의 생장과 연령 조성에 관한 연구를 수행하였고, 순간접착제를 이용한 곰피의 이식 등에 관한 연구(최 등 2002)가 보고된 바 있다. 그러나 지금까지 곰피의 식품 또는 약품제제로서의 기능성을 밝히는 연구는 아직 미미한 실정으로 이에 대한 체계적인 연구가 부족한 실정이다. 따라서 본 연구는 곰피추출물에 풍부하게 함유된 fucoidan, L-Ascorbic 산 및 collagen 등에 주목하여 곰피의 피부노화방지제로서의 가능성을 체계적으로 연구하기 위한 일환으로 우선 곰피추출물이 피부재생, 주름살제거 및 피부미백작용에 효과를 나타내는지 확인하기 위해 CCD-986sk cell line 및 Clone M-3 mouse melanocyte cell line을 이용하여 in vitro 수준에서 피부세포 생리활성 효과와 melanin formation 저해효과를 측정하였다. 그 결과 CCD-986sk cell line에 대한 세포생리활성 효과 실험에서 시험군인 곰피추출물 투여군은 대조군에 비해 강한 세포생리활성 효과를 나타내어 유의성 있는 실험결과를 보여주었고, Clone M-3 mouse melanocyte cell에 대한 melanin pigment 저해활성 효과 실험에서도 곰피추출물은 대조군에 비해 melanin pigment 생합성을 유의성 있게 감소시키는 실험결과를 나타내어 매우 흥미로운 결과를 보여주었다. 여러 가지 보고에 의하면 그동안 미국, 영국, 일본 등 구미 선진국의 과학자들은 intrinsic problems (피부구조 및 생리기능의 노쇠)과 extrinsic problems (자외선노출 및 스트레스)을 skin aging의 근본원인으로 규정짓고 피부 콜라겐과 엘라스틴을 재생시켜 피부 horny layer의 주름을 제거시키는 수 많은 종류의 chemical drugs를 개발하였고 현재도 개발 중에 있다 (Lavker et al. 1987; Choi and Oh 1997; Han et al. 1998). 그러나 이런 chemical에 비해 생체에 대한 부작용이나 세포독성이 상대적으로 적은 천연물을 이용하여 anti-skin aging drug을 개발하는 연구는 그 동안 부족했던 것이 현실이다. 한국은 오랜 역사를 통하여 화학물질이 아닌 다수의 천연물들을 임상에 적용하여온 역사를 가지고 있고 따라서 본 연구팀은 우리 남해안의 청정해역에서 자라는 곰피의 항피부 노화방지제로서의 개발 가능성에 주목하여 본 연구를 진행하였다. 연구 결과, 곰피추출물은 피부노화 및 skindarkness에 대한 대체치료물질 (alternative therapeutic agent)로서의 충분한 가능성을 보여주었다.

• Nutritional Sciences
• /
• 제8권4호
• /
• pp.242-249
• /
• 2005

It is well established that zinc plays an important role in bone metabolism and mineralization. The role of zinc in bone formation is well documented in animal models, but not much reported in cell models. In the present study, we evaluated zinc deficiency effects on osteoblastic cell proliferation, alkaline phosphatase activity and expression, and extracellular matrix bone nodule formation and bone-related gene expression in osteoblastic MC3T3-E1 cells. To deplete cellular zinc, chelexed-FBS and interpermeable zinc chelator TPEN were used. MC3T3-E1 cells were cultured in zinc concentration-dependent (0-15 ${\mu}M\;ZnCl_2$) and time-dependent (0-20 days) manners. MC3T3-E1 cell proliferation by MTT assay was increased as medium zinc level increased (p<0.05). Cellular Ca level and alkaline phosphatase activity were increased as medium zinc level increased (p<0.05). Alkaline phosphatase expression, a marker of commitment to the osteoblast lineage, measured by alkaline phosphatase staining was increased as medium zinc level increased. Extracellular calcium deposits measured by von Kossa staining for nodule formation also appeared higher in Zn+(15 ${\mu}M\;ZnCl_2$) than in Zn-(0 ${\mu}M\;ZnCl_2$). Bone formation marker genes, alkaline phosphatase and osteocalcin, were also expressed higher in Zn+ than in Zn-. The current work supports the beneficial effect of zinc on bone mineralization and bone-related gene expression. The results also promote further study as to the molecular mechanism of zinc deficiency for bone formation and thus facilitate to design preventive strategies for zinc-deficient bone diseases.

• 미생물학회지
• /
• 제50권3호
• /
• pp.173-178
• /
• 2014

Chondromyces crocatus의 자실체 형성에 대한 정량적 연구를 위하여 필수적인 분산된 세포현탁액 제조방법을 개발하였다. C. crocatus 세포들은 액체에서 서로 응집하는 특성이 있어 정량적 연구가 어렵다. 하지만 3% 한천을 함유한 casitone-yeast extract 평판배지에서 배양한 세포들은 물에 잘 분산되어 분산된 세포현탁액을 제조할 수 있었다. 이렇게 제조한 $2{\times}10^8cells/ml$ 농도의 세포현탁액 $20{\mu}l$를 영양분이 고갈된 평판배지 위에 올려놓고 배양하였을 때 C. crocatus 특유의 자실체를 형성하였다. Casitone과 같은 영양분을 첨가하였을 때는 자실체 형성이 저해되거나 변형되었으며, 한천의 농도가 높을수록 자실체의 가지수가 증가되었다. 조사된 최적조건에서 C. crocatus KYC2823은 24시간 내로 자실체 구조를 완성하였다.

• Archives of Pharmacal Research
• /
• 제28권11호
• /
• pp.1311-1316
• /
• 2005

To better define the relationship between dermal regeneration and wound contraction and scar formation, the effects of epidermal growth factor (EGF) loaded in collagen sponge matrix on the fibroblast cell proliferation rate and the dermal mechanical strength were investigated. Collagen sponges with acid-soluble fraction of pig skin were prepared and incorporated with EGF at 0, 4, and 8 $\mu$g/1.7 $cm^{2}$. Dermal fibroblasts were cultured to 80$\%$ confluence using DMEM, treated with the samples submerged, and the cell viability was estimated using MTT assay. A deep, $2^{nd}$ degree- burn of diameter 1 cm was prepared on the rabbit ear and the tested dressings were applied twice during the 15-day, post burn period. The processes of re-epithelialization and dermal regeneration were investigated until the complete wound closure day and histological analysis was performed with H-E staining. EGF increased the fibroblast cell proliferation rate. The histology showed well developed, weave-like collagen bundles and fibroblasts in EGF-treated wounds while open wounds showed irregular collagen bundles and impaired fibroblast growth. The breaking strength (944.1 $\pm$ 35.6 vs. 411.5 $\pm$ 57.0 Fmax, $gmm^{-2}$) and skin resilience (11.3 $\pm$ 1.4 vs. 6.5 $\pm$ 0.6 mJ/$mm^{2}$) were significantly increased with EGF­treated wounds as compared with open wounds, suggesting that EGF enhanced the dermal matrix formation and improved the wound mechanical strength. In conclusion, EGF-improved dermal matrix formation is related with a lower wound contraction rate. The impaired dermal regeneration observed in the open wounds could contribute to the formation of wound contraction and scar tissue development. An extraneous supply of EGF in the collagen dressing on deep, $2^{nd}$ degree-burns enhanced the dermal matrix formation.

• Journal of Microbiology and Biotechnology
• /
• 제25권8호
• /
• pp.1390-1400
• /
• 2015

Quorum sensing is a process of cell-to-cell communication in which bacteria produce autoinducers as signaling molecules to sense cell density and coordinate gene expression. In the present study, a LuxI-type synthase, AnoI, and a LuxR-type regulator, AnoR, were identified in Acinetobacter nosocomialis, an important nosocomial pathogen, by sequence analysis of the bacterial genome. We found that N-(3-hydroxy-dodecanoyl)- _L -homoserine lactone (OH-dDHL) is a quorum-sensing signal in A. nosocomialis. The anoI gene deletion was responsible for the impairment in the production of OH-dDHL. The expression of anoI was almost abolished in the anoR mutant. These results indicate that AnoI is essential for the production of OH-dDHL in A. nosocomialis, and its expression is positively regulated by AnoR. Moreover, the anoR mutant exhibited deficiency in biofilm formation. In particular, motility of the anoR mutant was consistently and significantly abolished compared with that of the wild type. The deficiency in the biofilm formation and motility of the anoR mutant was significantly restored by a functional anoR, indicating that AnoR plays important roles in the biofilm formation and motility. Furthermore, the present study showed that virstatin exerts its effects on the reduction of biofilm formation and motility by inhibiting the expression of anoR. Consequently, the combined results suggest that AnoIR is a quorum-sensing system that plays important roles in the biofilm formation and motility of A. nosocomialis, and virstatin is an inhibitor of the expression of anoR.

• 한국자기학회:학술대회 개요집
• /
• 한국자기학회 2014년도 임시총회 및 하계학술연구발표회
• /
• pp.120-121
• /
• 2014

We investigate the field-dependence of energy barrier for various cell diameters and two type of geometry through the NEB method. We find that the energy barrier can depend strongly on the cell size when the switching is governed by the domain wall motion. Moreover we also examine the cell size dependence of energy barrier for two type of cell geometry. In the presentation, we will discuss the effect of domain wall formation and more various cell size on the energy barrier in detail.

• Journal of Microbiology and Biotechnology
• /
• 제4권3호
• /
• pp.191-194
• /
• 1994

Acetoin and ammonia, the precursors of tetramethylpyrazine (TMP) having "nutty " or "roasted" flavors, were produced by cultivating Lactococcus lactis ssp. lactis biovar. diacetilactis FC1. The effects of the volume concentration ratio (VCR) of cell-free medium on the formation of TMP were investigated using a rotary evaporator at $70^{\circ}C than at 80^{\circ}C$. As the VCR increased, the formation of TMP and the conversion ratio of acetoin to TMP increased. More TMP were formed at $70^{\circ}C than at 80^{\circ}C$. As the VCR increased, the concentration of acetoin decreased implying the formation of TMP from acetoin and ammonia.

• Bulletin of the Korean Chemical Society
• /
• 제32권3호
• /
• pp.909-914
• /
• 2011

An improved convergent and economical method has been developed for the synthesis of erlotinib, a 4-anilinoquinazoline and an EGFR-tyrosine kinase inhibitor for treatment of non-small-cell lung cancer. The final two steps for the formation of this 4-anilinoquinazoline from suitable 2-aminobenzonitrile intermediate and 3-ethynylaniline were modified and were performed in a simple one-pot reaction. The ring-closing mechanism for the formation of erlotinib from the suitable formamidine intermediate and 3-ethynylaniline was investigated and determined to proceed via the formation of phenyl benzamidine intermediate rather than involving Dimroth rearrangement reported earlier. The new benzamidine intermediate was isolated for the first time and characterized.