The role of CXCR7, a seven-transmembrane G-protein coupled chemokine receptor, which binds with high affinity to chemokine CXCL11 and CXCL12 in oral cancer cells and the effect of transient CXCR7-downregulation on proliferation and migration of oral squamous cell carcinoma (OSCC) cells have not been reported. The aim of the present study was to evaluate the effects of CXCR7 on an OSCC cell line. In this study, we down-regulated CXCR7 in the KOSCC25B OSCC cell line by siRNA. In vitro cell proliferation and migration assays were used to investigate the effect of CXCR7- downregulation on cell proliferation and migration in si.KOSCC25B cells. The CXCR7 down-regulated OSCC cells grew significantly slower than the negative control siRNA transfected KOSCC25B cells (p<0.05). Additionally, migration of si.KOSCC25B cells decreased significantly compared with non-transfected KOSCC25B cells (p<0.007). These results suggest that down-regulation of CXCR7 induces anti-proliferative and anti-migratory effects in OSCC, and that CXCR7 may be a useful target molecule for the treatment of OSCC.
An efficient method for in vitro propagation and growth of the wild garlic(Allium ochotense Prokh.) was established. Bulbs of wild garlic were collected from Ullung Island, Korea, and the growth pattern of plantlets on various culture media was observed. High growth of shoot was obtained on LP, NN and N6 medium. The growth media supplemented with 3%(w/v) sucrose was found optimal for shoot growth. After 10 weeks multiple shoots were observed in the plantlets growing on the medium containing 1.0 mg/l of zeatine and 0.1 mg/l NAA. Roots were induced directly at the base of the shoot in all treatments. The medium with 2.0 mg/l of IBA proved to be the best rooting medium. The studies of this kind may be used to develop strategies for large-scale propagation of elite wild garlic.
Heterodera trifolii, mostly known as clover cyst nematode, is currently a serious problem for Chinese cabbage growers of the highland area in Korea. Due to lack of readily information about the nematode on Chinese cabbage in Korea, the pest steadily spread within the highland areas and has become a serious setback. Occurrence, spatial aggregation, egg hatching and the pathogenicity of this nematode are depicted in this study from ecological point of view. The study results suggest site-specific control and a potential planting time for the cabbage to avoid severe damage caused by this nematode.
In the present study, we investigated the expression patterns of p63, a member of the p53 gene family, in hair follicle cells at different stages of the hair cycle and examined the relation with cell proliferation activity. For this study, immunohistochemistry for p63 and Ki-67, a marker of cell proliferation, was performed in skin obtained from C3H/he mice with depilation. In the anagen stage, p63 was strongly expressed in the cells of bulge areas and epithelial strand, matrix cells of the hair bulbs and outer root sheath cells, but inner root sheath cells and dermal papilla cells were negative for p63. These expression patterns of p63 were similarly noted in hair follicles in the early catagen stage. In the late catagen and telogen stages of hair follicles, outer root sheath cells, seboblasts and duct cells were immunoreactive for p63. On the other hand, Ki-67-positive cells were selectively observed among the p63 positive cell components, although p63 positive cells were not always proliferative. Most of the matrix cells in the hair bulbs were positive for Ki-67. Ki-67-positive cells were also frequently evident in the cells of epithelial strands in the early anagen stage. Outer root sheath cells were often positive for Ki-67 in the anagen and early catagen stages, but very rare in the late catagen and telogen stages. In summary, p63 was expressed in the bulge stem cells, epithelial strand cells, matrix cells and outer root sheath cells of hair follicles at any stage of the cycle, which was associated with the movement of hair progenitor cells for regeneration. Ki-67-positive cells were evident among the p63-expressing cell components. Our results strongly suggest that p63 plays an important role in stem cell regulation, at least associated with cell proliferation, for the regeneration of hair follicles.
이 연구는 제주연안 해역에서 갯녹음의 확산동향과 기후변화로 인한 동계 수온변화(수온상승)가 갯녹음 확산에 미치는 영향을 알아보고 갯녹음 원인생물인 무절석회조류의 번식과 생장을 파악하기 위해 연구되었다. 제주연안의 갯녹음 발생면적은 1998년에는 2,931ha였으나, 2003년에는 4,541ha로 증가하였다. 발생해역도 1998년은 제주도 남부 해역에서 주로 발생했으나, 2003년에는 조천읍, 구좌읍을 제외한 제주도 전역으로 확산되었다. 1992년부터 2004년까지 관측된 2월 평균수온은 갯녹음 해역 15.1℃, 해중림 해역 13.9℃로 갯녹음 해역이 1.2℃가 높게 나타나 두 해역 간 뚜렷한 차이를 보였으나, 8월 수온은 두 해역 간 차이가 없었다. 수온의 장기변동(37년)에서도 갯녹음 해역이 평균 15.3℃인데 비하여 해중림 해역은 평균 14.1℃로 갯녹음 해역이 해중림 해역보다 1.2℃가 높게 나타났다. 연간 수온 증가 값은 갯녹음 해역이 매년 0.038℃씩 증가하고 있는 반면, 해중림 해역은 0.024℃씩 증가하여 장기 수온변동은 갯녹음 해역이 해중림 해역보다 높았다. 이와 같이 기후변화로 인한 지속적인 동계 수온상승은 제주도 갯녹음을 확대시 키고 있음을 시사하고 있다.
As diethylnitrosamine (DEN) effect on cell proliferation, DNA damage and stem cell marker(s) expression have been largely unknown in mouse normal hepatocytes (AML-12 cells) cultured over a short-term period, this study was conducted to examine the cell proliferation, Ataxia telangiectasia mutated (ATM) and epithelial cell adhesion molecule (EpCAM) and Neighbor of Punc E 11 (Nope) expression in AML-12 cells treated with DEN for 24 and 48 h. Cells were treated with DEN (25-800 μg/mL) and cell phenotype was determined, and the MTT assay was used to quantify the proliferation of cells treated with DEN. Expression and distribution of ATM in AML-12 cells were determined by indirect immunofluorescence microscopy. And Western blot analysis of EpCAM and Nope was performed. Cell viability was significantly increased in response to all doses of DEN treatment compared to control at 24 h (p<0.05 or p<0.01). However, there was no significant increase at 48 h, even though it showed increased trend. Immunofluorescence staining of ATM showed that there was an increase of ATM expression at doses of 50, 100 and 200 μg/mL of DEN treatment, showing strong nuclear staining. Furthermore, Western blot analysis showed that DEN treatment showed increased trend of EpCAM and Nope expression. Taken together, DEN treatment increased cell proliferation in AML- 12 cells, and it was associated with increased ATM expression.
Coffee is one of the most familiar beverages to modern human adults, but its bio-physiological effect has not been clearly elucidated. It was known that more than one thousand chemicals were included in the ordinary coffee extract. Among them, the caffein and chlorogenic acid (caffeoylquinic acids) are most abundant and have been investigated by many authors so far. In order to know the real cellular effect of whole coffee extract elements, the dialyzed coffee extract (DCE)1) was made to get coffee elements less than 1000 Da molecular weight, which are freely absorable through gastrointestinal tract. It was directly treated in the culture of RAW 264.7 cells, a murine macrophage lineage. RAW 264.7 cells were treated with DCE equivalent to 2.5 cups of coffee (DCE-2.5), DCE-5, and DCE-10 for 12 hours, and their protein extracts were examined by histological observation and immunoprecipitation high performance liquid chromatography (IP-HPLC). RAW 264.7 cells differently expressed the proliferation-related proteins depending on the dose of DCE. DCE-2.5 and DCE-5 enhanced the cellular growth of RAW 264.7 cells by increasing the expression of β-actin, PCNA, Ki-67, MPM2, MAX, cMyc, E2F-1, and Rb-1, and by decreasing the expression of MAD and p21. These proliferation-related proteins were rarely affected by DCE-10. DCE-2.5 and DCE-5 induced the cellular proliferation of RAW 264.7 cells by the signaling of E2F-1 and cMyc, respectively, but these cellular effects almost disappeared in DCE-10. Therefore, it was presumed that the low dose of coffee, DCE-2.5 and DCE-5 might be effective for the proliferation of murine macrophages, RAW264.7 cells, contrast to the high dose of coffee, DCE-10. It was also suggested that the low dose of DCE-2.5 and DCE-5 be helpful to increase the innate immunity in vivo by increasing the cell number of macrophages in contrast to the high dose of DCE-10.
To investigated the mechanism, induced pluripotent stem cells(iPSC) is important for clinical application and stem cell research. It is well known that hMAGEA2 expression pattern and effect on differentiation in embryonic stem cell but their specific role in iPS cells are unclear. The present study was schemed to understand the function of hMAGEA2 gene in iPS cells and to elucidate its characteristic. Although overexpression of hMAGEA2 in iPS cells are not different on morphology, their pluripotency and self-renewal capacity are significantly strengthened. And hMAGEA2 contributed to promote the cell cycle progression, this cell cycle changes induced proliferation acceleration. Through embryoid body formation in vitro and teratoma formation in vivo, we found that hMAGEA2 critically decreases the differentiation ability in iPS cells. Our results demonstrate that hMAGEA2 intensified the self-renewal, pluripotency, proliferation degree but efficiency of differentiaton is significantly repressed. Our findings provided that hMAGEA2 play a key role of iPS cells.
Aneurysmal bone cyst (ABC) in maxilla is a rare and benign lesion but shows extensive bony destruction, occasionally accompanied with secondary osseous lesions, i.e., central giant cell granuloma, ossifying fibroma, fibrous dysplasia, etc. As the pathogenesis of ABC has not been clearly defined, ABC is diagnostically challenged due to its variable histological features. A 17-year-old boy showed a huge radiolucent lesion at right anterior maxilla, which was accidentally found in routine dental-radiological examination for orthodontic treatment. He had no medical history of systemic disease, and did not remember any traumatic experience on his right anterior maxilla. The radiolucent lesion involved periapical area from right central incisor to right first premolar, and was clinically diagnosed as odontogenic keratocyst. During surgical operation a cyst-like sac was enucleated with severe hemorrhage. In the histological observation the thick fibrous sac showed no lining epithelium, and its luminal side disclosed multiple aneurysmal spaces which were shrunken and almost obliterated. The fibrous sac itself was hyperplastic with abundant vascular channels, and produced fibromatous thickening associated with ossifying trabecular bones. This fibro-osseous tissue was hamartomatous, which was not directly connected and organized with marrow bone of maxilla. Finally, the present case was diagnosed as secondary type ABC differentially from traumatic bone cyst (TBC), odontogenic cyst, and central reparative granuloma. And it was presumed that the hamartomatous proliferation of fibro-osseous tissue in the cystic sac of ABC could produce the swelling pressure effect in the bone marrow similar to the overgrowth of central giant cell granuloma, ossifying fibroma, fibrous dysplasia, etc., in the secondary type ABC.
해양생물을 포함한 천연물질은 신약개발의 원천 소재로서 매력적이며, 특히 무수한 미지의 해양생물들의 연구가 관심을 받고 있다. 기존의 연구에서 미크로네시아에서 채취한 해면동물 40여종에 대하여 항증식 효과를 다양한 암세포주에서 검색한 바 있다. 본 연구에서는 그 중 Cos-cinoderma sp.의 작용 및 그 기전을 살펴보았다. 특히, 암억제유전자 p53의 발현을 억제시킨 세포주(HCT116 p53KO 과 RKO-E6)에서의 차이점을 비교하였다. 세포생존률 시험에서 Coscinoderma sp. 추출물은 p53의 유무와 상관없이 암세포의 증식을 억제하였음을 확인하였다. 이 암세포증식 억제 효과가 p53 존재에 따라 다르게 나타나는지 알아보기 위하여 세포사멸 관련 단백질 발현양을 Coscinoderma sp. 처리한 각 세포주에서 비교하였다. 그 결과, Coscinoderma sp.를 HCT16 세포주에 처리하였을 때, p53과 Noxa의 발현이 증가하는 것을 관찰하였고, caspase-9이 분절되면서 감소하는 것으로부터 apoptosis를 일으킨다고 여겨진다. 반면, p53이 결핍된 HCT116세포주에서는 Coscinoderma sp. 에 의하여 p21과 mTOR의 발현이 증가되는 것을 확인하였고, 이는 senescence를 야기할 수 있다고 여겨진다. 본 연구로부터 Coscinoderma sp.는 p53의 존재여부에 따라 상이한 작용기전을 매개하여 대장암 세포주의 증식을 억제한다는 것을 알 수 있었다. 이는 새로운 항암제의 개발 가능성을 제시하는 것으로, Coscinoderma sp.의 활성 성분에 대한 지속적인 연구가 이루어 져야 할 것으로 보인다.
Skin-derived precursors (SKPs) have potential to differentiate to various cell types including osteoblasts, adipocytes and neurons. SKPs are a candidate for cell-based therapy since they are easily accessible and have multipotency. Most mammalian cells are exposed to a low oxygen environment with 1 to 5% O2 concentration in vivo, while 21% O2 concentration is common in in vitro culture. The difference between in vitro and in vivo O2 concentration may affect to the behavior of cultured cells. In this report, we investigated the effect of hypoxic condition on stemness and proliferation of SKPs. The results indicated that SKPs exposed to hypoxic condition for 5 days showed no change in proliferation. In terms of mRNA expression, hypoxia maintained expression of stemness markers; whereas, oncogenes, such as Klf4 and c-Myc, were downregulated, and the expression of Nestin, related to cancer migration, was also downregulated. Thus, SKPs cultured in hypoxia may reduce the risk of cancer in SKP cell-based therapy.
The PHBV nonofibrous membrane fabricated by electron-spinning method for tissue engineered bone regeneration scaffold was evaulated in terms of cellular prolieration and cryopreservation efficiency. The rat calvarial periosteum derived primary cells were cultured with PHBV nanofibrous membranes and analyzed the cellular proliferation and differention fashion and cryopreservation potential by in vitro MTT assay as well as ALP staining and Alizarine red staining with or without cryopreservation for 2 weeks. The rat calvarial periosteum derived primary cells cultured with PHBV nonofibrous membrane showed favorable proliferation and alkaline phosphatase activity with numerous mineral nodule formation regardless of cryopreservation, even though its efficiency was slightly decreased in cryopreserved condition. These findings suggest that PHBV nanofibrous membrane can be applicable as an efficient cell engineered membrane for guided bone regeneration or scaffold for tissue engineered bone regeneration.