Background: The calcium-binding S100A4 protein is involved in epithelial to mesenchymal transition, oncogenic transformation, angiogenesis, cytoskeletal integrity, mobility and metastasis of cancer cells. This study aimed to clarify the roles of S100A4 in genesis and progression of glioma. Materials and Methods: S100A4 expression was examined by real-time RT-CPR and Western blot in glioma and paired normal brain tissue (n=69), and compared with clinicopathological parameters of tumors. In addition, glioma U251 cells transfected with an S100A4-expressing plasmid were examined for proliferation by MTT, apoptosis by Annexin V-FITC, and migration and invasion with Transwell chambers. Results: Increased S100A4 mRNA expression was found in gliomas, compared with paired non-tumor tissue (p<0.001). Gradual elevation of overexpression of S100A4 was observed with increasing glioma grade (p<0.001). Astrocytoma showed lower S100A4 mRNA expression than oligodendrogliomas, with glioblastomas having highest values (p<0.001). Similar results were obtained for S100A4 protein, a positive link being found between mRNA and protein expression in gliomas (p<0.001). There was higher growth, lower apoptosis, stronger migration and invasion of S100A4 transfectants than control and mock transfected cells (p<0.001). Conclusions: These findings indicate that up-regulated S100A4 expression is positively linked to pathogenesis, progression and histogenesis of glioma by modulating proliferation, apoptosis, migration and invasion.
We report a method for optical monitoring of tumors in an animal model using optical coherence tomography (OCT). In a spectral domain OCT system, a superluminescent diode light source with a full width of 66 nm at half maximum and peak wavelength of 950 nm was used to take images having an axial resolution of 6.8 ${\mu}m$. Cancer cells of PC-3 were cultured and inoculated into the hypodermis of auricle tissues in BALB/c nude mice. We observed tumor formation and growth at the injection region of cancer cells in vivo and obtained the images of tumor mass center and sparse circumferences. On the $5^{th}$ day from an inoculation of cancer cells, histological images of the tumor region using cross-sectional slicing and dye staining of specimens were taken in order to confirm the correlation with the high resolution OCT images. The OCT image of tumor mass compared with normal tissues was analyzed using its A-scan data so as to obtain a tissue attenuation rate which increases according to tumor growth.
The aim of this study was to evaluate gamma-aminobutyric acid (GABA)-induced erythropoietin (EPO) and EPO-receptor expression in human Hep3B cells and Sprague Dawley (SD) rats during hypoxia. Expression levels of EPO, EPO-R mRNA, Janus kinase-2 (JAK-2), vascular endothelial growth factor (VEGF), hypoxia inducible factor-1 (HIF-1), and HIF-2 in response to GABA treatment were evaluated in cell lines. SD rats were randomly divided into 5 groups of 8 rats each, and GABA was orally administered; the groups were the normal control (NC), hypoxia-exposed (G0), as well as the GABA 1 mg/100 g body weight (BW) GABA treated group (G1), 5 mg/100 g BW GABA treated group (G5), and 10 mg/100 g BW GABA treated group (G10) with hypoxia. We analyzed EPO levels and red blood cell counts in rat blood and EPO gene expression in kidney tissue. EPO and VEGF mRNA levels in Hep3B cells exposed to hypoxia were significantly increased and further increased after GABA treatment. However, the expression of EPO-R and JAK-2 mRNAs were not affected by GABA, but hypoxia-induced HIF-1 and HIF-2 mRNA expression was inhibited by GABA. In the kidney tissue of rats exposed to hypoxia, the expression level of EPO mRNA was greatly increased, but levels in the GABA treatment groups significantly decreased. EPO levels in the serum showed the same significant trend, but the red blood cell counts were not significantly different. These findings demonstrate that HIF-1 and HIF-2 activation increase EPO expression in Hep3B cells exposed to hypoxia. However HIF decreased by GABA addition and VEGF increased significantly.
버들치 Rhynchocypris oxycephalus의 등지느러미와 가슴지느러미를 연결하는 중간 부위 체측상처를 수술하여, 과립형성 조직 수축 특성과 치유율 및 2차 치유에 관하여 조사하였다. 수술 푸 생존율은 수술 직후 및 수술 후 7일에 대조군의 100%에 비해 각각 95%와 99% 이었다. 수술 후 14일과 42일 사이에 사망은 없었다. 수술후 7~21에 과립형성 조직은 상피화되었으며 수술 후 29일에 과립형성 조직은 상처 주변부로 정상 위치수축을 하였다. 수술 후 35일~42일에 과립형성 조직에는 collagen 섬유가 존재하였고 표피는 잘 분화된 , alarm 물질세포를 내포하고 있었으며 상처 부위의 표피는 모든 어류에서 조직학적으로 정사이어서, 상처 치유는 수술 35일 이내에 완전히 이루어졌다.
Objectives: This study was performed to investigate the effects of Hominis Placenta pharmacopuncture (HPP) therapy on the experimentally-induced endometriosis in the rats. Materials and Methods: Endometriosis was induced in rats by autotransplanting uterine tissue to the peritoneum and divided them into three groups: (1) sham-operated group (n=8), (2) surgically induced endometriosis and untreated control group (n=8), (3) surgically induced endometriosis and HPP treated group. Sham-operated group and control group were inject with normal saline once a every other day for 30days, while treated group was injected with HPP extract once a every other day for same duration. Injected point of HPP and normal saline were subcutaneous tissue at Gwanwon (CV4) acupoint. Then we measured the body weight, the volume of endometriotic implants, the weigh of uterus and ovaries, and investigated the concentration of cytokines (MCP-1, TNF-${\alpha}$) in peritoneal fluids. Histopathology, immunohistochemisty for COX-2 and VEGF, and histochemistry for mast cell in transplanted uterine tissue were performed. Results: The volume ($mm^2$) of endometriotic implants in HPP treated group ($55.4{\pm}41.6$) was significantly decreased (p<0.01) compared with control group ($140{\pm}66.1$). And the concentration (pg/ml) of MCP-1 in peritoneal fluids in HPP treated group ($1117.6{\pm}60.5$) was significantly decreased (p<0.01) compared with control group ($1446.2{\pm}280.3$). The concentration (pg/ml) of TNF-${\alpha}$ in peritoneal fluids in HPP treated group ($80.6{\pm}31.4$) was decreased (p<0.01) compared with control group ($145.3{\pm}86.9$). Histopathologically, proliferation of endometriotic epithelia, infiltration of inflammatory cell and angiogenesis in transplanted uterine tissue of HPP treated group were weakly observed than those of control group. The COX-2 expression in endometrial, epithelial and stromal cells in transplanted uterine tissue of HPP treated group was decreased compared with control group. The VEGF expression of endometriotic epithelia, neovascular endothelia and stromal cell in transplanted uterine tissue of HPP treated group were weakly observed than those of control goup. Conclusions: HPP is effect on Endometriosis of rats by Experimentally-induced.
Journal of the Korean Association of Oral and Maxillofacial Surgeons
/
제34권3호
/
pp.245-249
/
2008
Epidermal growth factor is a single-chain polypeptide consisting of 53 amino acids and has a potent mitogenic activity that stimulates proliferation of various normal and neoplastic cells through the interaction with its specific receptor(epidermal growth factor receptor, EGFR). Pleomorphic adenoma is the most common salivary benign tumor and histologically, it contains the epithelial cell, the myo-epithelial cell and mesenchymal ingredient, which is various aspect. Adenoid cystic carcinoma is an infiltrative malignant salivary gland tumor with three different histological patterns: cribriform, tubular or solid. The tumor cell structure composed of modified myoepithelial cell, and basaloid cell. In this study, we used an immunohistochemical technique to investigate the expression of EGF in 6 specimens of adenoid cystic carcinoma and 10 specimens of pleomorphic adenoma taken from patients treated at Dept. of Oral and Maxillofacial Surgery, Dankook University. The results were as follows. 1. In pleomorphic adenoma, ductal structure and scattered spindle cells in hyalinized stroma, disclosing myxoid stroma and hyalin, cartilage formation were observed. Immunohistologically, weak EGF expression in ductal structure and negative in stromal area were observed. 2. Cribriform type of adenoid cystic carcinoma showed numerous pseudocyst surrounded by dark small neoplastic cells in the back-ground of fibrous connective tissue and moderate EGF expression of dark cells adjacent to pseudo lumen in cribriform pattern, while weak expression in other most cells. 3. Tubular type of adenoid cystic carcinoma showed numerous ductal pattern surrounded by two layered neoplastic cells in the back-ground of fibrous connective tissue and strong EGF expression in luminal cells of ductal structure, while weak expression in outer cells. From the results obtained, we suggest that EGF is mainly biosynthesized in cells forming duct like structures of tubulo-ductal type or cribriform adenoid cystic carcinoma and it may play a role, as a cell mitogen in adenoid cystic carcinoma growth.
Periodontal ligament(PDL) cells and human gingival fibroblasts(HGFs) play important roles in development, regeneration, normal function, and pathologic alteration. PDL cells and HGFs have the similarity related with general characteristics of fibroblast such as spindle shaped morphology, the presence of vimentin intermediate filament and the synthesis of interstitial collagens and fibronectin. There were many studies about the differences between PDL cells and HGFs, but they were not about whole gene level. In this study, we tried to explain the differences of gene expression profiles between PDL cells and HGFs, and the differences among three individuals by screening gene expression patterns of PDL cells and HGFs, using cDNA microarray. Although there were some variants among three experiments, a set of genes were consistentely and differentially expressed in one cell type. Among 3,063 genes, 49 genes were more highly expressed in PDL cells and 12 genes were more highly expressed in HGFs. The genes related with cell structure and motility were expressed more highly in PDL cells. These are cofilin 1, proteoglycan 1 secretory granule, collagen type I(${\alpha}$ 1), adducin gamma subunit, collagen type III(${\alpha}$ 1), fibronectin, lumican(keratan sulfate proteoglycan), and ${\alpha}$ -smooth muscle actin. Tissue inhibitor of metalloproteinase known as the enzyme controlling extracellular matrix with matrix metalloproteinase is more highly expressed in PDL cells, osteoprotegerin known as osteoclastogenesis inhibitory factor is more highly expressed in HGFs. We performed northern blot to verify cDNA microarray results on selected genes such as tissue inhibitor of metalloproteinase, fibronectin, osteoprogeterin. The result of northern blot analysis showed that each cell expressed the genes in similar pattern with cDNA microarray result. This result indicates that cDNA microarray is a reliable method in screening of gene expression profiles.
Purpose: Hrd1 has recently emerged as a critical regulator of B-cells in autoimmune diseases. However, its role in the pathogenesis of chronic rhinosinusitis with nasal polyps (CRSwNP) remains largely unexplored. This study aimed to examine Hrd1 expression and B-cell accumulation and their possible roles in CRSwNP. Methods: Quantitative real-time polymerase chain reaction, immunohistochemistry, enzyme-linked immunosorbent assay and Western blotting were used to assess gene and protein expression in nasal tissue extracts. Cells isolated from nasal tissues and peripheral blood mononuclear cells were characterized by flow cytometry. Local antibody production was measured in tissue extracts with a Bio-Plex assay. Additionally, changes in Hrd1 expression in response to specific inflammatory stimuli were measured in cultured dispersed polyp cells. Results: Nasal polyps (NPs) from patients with eosinophilic CRSwNP (ECRS) had increased levels of Hrd1, B-cells and plasma cells compared with NPs from patients with non-eosinophilic CRSwNP (non-ECRS) or other control subjects (P < 0.05). The average Hrd1 levels in B-cells in NPs from ECRS patients were significantly higher than those from non-ECRS patients and control subjects (P < 0.05). NPs also contained significantly increased levels of several antibody isotypes compared with normal controls (P < 0.05). Interestingly, Hrd1 expression in cultured polyp cells from ECRS patients, but not non-ECRS patients, was significantly increased by interleukin-$1{\beta}$, lipopolysaccharide and Poly(I:C) stimulation, and inhibited by dexamethasone treatment (P < 0.05). Conclusions: Differential Hrd1 expression and B-cell accumulation between the ECRS and non-ECRS subsets suggests that they can exhibit distinct pathogenic mechanisms and play important roles in NP.
In 1997 when cloned sheep Dolly and soon after Polly were born, it had become head-line news because in the former the nucleus that gave rise to the lamb came from cells of six-year-old adult sheep and in the latter case a foreign gene was inserted into the donor nucleus to make the cloned sheep produce human protein, factor IX, in e milk. In the last few years, once the realm of science fiction, cloned mammals especially in livestock have become almost commonplace. What the press accounts often fail to convey, however, is that behind every success lie hundreds of failures. Many of the nuclear-transferred egg cells fail to undergo normal cell divisions. Even when an embryo does successfully implant in the womb, pregnancy often ends in miscarriage. A significant fraction of the animals that are born die shortly after birth and some of those that survived have serious developmental abnormalities. Efficiency remains at less than one % out of some hundred attempts to clone an animal. These facts show that something is fundamentally wrong and enormous hurdles must be overcome before cloning becomes practical. Cloning researchers now tent to put aside their effort to create live animals in order to probe the fundamental questions on cell biology including stem cells, the questions of whether the hereditary material in the nucleus of each cell remains intact throughout development, and how transferred nucleus is reprogrammed exactly like the zygotic nucleus. Stem cells are defined as those cells which can divide to produce a daughter cell like themselves (self-renewal) as well as a daughter cell that will give rise to specific differentiated cells (cell-differentiation). Multicellular organisms are formed from a single totipotent stem cell commonly called fertilized egg or zygote. As this cell and its progeny undergo cell divisions the potency of the stem cells in each tissue and organ become gradually restricted in the order of totipotent, pluripotent, and multipotent. The differentiation potential of multipotent stem cells in each tissue has been thought to be limited to cell lineages present in the organ from which they were derived. Recent studies, however, revealed that multipotent stem cells derived from adult tissues have much wider differentiation potential than was previously thought. These cells can differentiate into developmentally unrelated cell types, such as nerve stem cell into blood cells or muscle stem cell into brain cells. Neural stem cells isolated from the adult forebrain were recently shown to be capable of repopulating the hematopoietic system and produce blood cells in irradiated condition. In plants although the term$\boxDr$ stem cell$\boxUl$is not used, some cells in the second layer of tunica at the apical meristem of shoot, some nucellar cells surrounding the embryo sac, and initial cells of adventive buds are considered to be equivalent to the totipotent stem cells of mammals. The telomere ends of linear eukaryotic chromosomes cannot be replicated because the RNA primer at the end of a completed lagging strand cannot be replaced with DNA, causing 5' end gap. A chromosome would be shortened by the length of RNA primer with every cycle of DNA replication and cell division. Essential genes located near the ends of chromosomes would inevitably be deleted by end-shortening, thereby killing the descendants of the original cells. Telomeric DNA has an unusual sequence consisting of up to 1,000 or more tandem repeat of a simple sequence. For example, chromosome of mammal including human has the repeating telomeric sequence of TTAGGG and that of higher plant is TTTAGGG. This non-genic tandem repeat prevents the death of cell despite the continued shortening of chromosome length. In contrast with the somatic cells germ line cells have the mechanism to fill-up the 5' end gap of telomere, thus maintaining the original length of chromosome. Cem line cells exhibit active enzyme telomerase which functions to maintain the stable length of telomere. Some of the cloned animals are reported prematurely getting old. It has to be ascertained whether the multipotent stem cells in the tissues of adult mammals have the original telomeres or shortened telomeres.
The Bric-a-brac, Tramtrack, Broad-complex (BTB) domain is a protein-protein interaction domain that is found in many zinc finger transcription factors. BTB containing proteins play important roles in a variety of cellular functions including regulation of transcription, regulation of the cytoskeleton, protein ubiquitination, angiogenesis, and apoptosis. Here, we report the cloning and characterization of a novel human gene, KLHL31, from a human embryonic heart cDNA library. The cDNA of KLHL31 is 5743 bp long, encoding a protein product of 634 amino acids containing a BTB domain. The protein is highly conserved across different species. Western blot analysis indicates that the KLHL31 protein is abundantly expressed in both embryonic skeletal and heart tissue. In COS-7 cells, KLHL31 proteins are localized to both the nucleus and the cytoplasm. In primary cultures of nascent mouse cardiomyocytes, the majority of endogenous KLHL31 proteins are localized to the cytoplasm. KLHL31 acts as a transcription repressor when fused to GAL4 DNA-binding domain and deletion analysis indicates that the BTB domain is the main region responsible for this repression. Overexpression of KLHL31 in COS-7 cells inhibits the transcriptional activities of both the TPA-response element (TRE) and serum response element (SRE). KLHL31 also significantly reduces JNK activation leading to decreased phosphorylation and protein levels of the JNK target c-Jun in both COS-7 and Hela cells. These results suggest that KLHL31 protein may act as a new transcriptional repressor in MAPK/JNK signaling pathway to regulate cellular functions.
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