Imperatorin, a biologically active furanocoumarin from the roots of Angelica dahurica (Umbelliferae), was mutagenic and induced transformation of mouse fibroblast cell lines, whereas it provided inhibiting effects on mutagenesis and carcinogenesis induced by various carcinogens. Furthermore, it has been suggested that imperatorin may have potential anticarcinogenic effects when administered orally in the diet. In addition to its anticarcinogenic properties, imperatorin has been shown to possess anticancer activities. We investigated the macro scale gene expression analysis on the HL-60 cells treated with imperatorin. Imperatorin (10μM) were used to treat the cells for 6h, 12h, 24h, 48h, and 72h. In a human cDNAchip study of 10,000 genes evaluated 6, 12, 24, 48, 72 hours after treated with imperatorin in HL-60 cells. Hierarchical cluster against the genes which showed expression changes by more than 2 fold. Three hundred eighty six genes were grouped into 6 clusters by a hierarchical clustering algorithm. Pathway analysis using gene microarray pathway prof Her that is a computer application designed to visualize gene expression data on screen representing biological pathways and groupings of genes.
Journal of the Korean Association of Oral and Maxillofacial Surgeons
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제30권4호
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pp.261-270
/
2004
The purpose of this experiment was to examine the histological changes and the pattern of expression of type I, II collagen in the elongated area by distraction osteogenesis in the rabbit mandible. Sixteen rabbits weighing 2.5kg-3kg were used for this experiment. Experimental group was distracted at the rate of 0.7mm, twice/day for 7days, and control group was only osteotomized. After 5 days latency, osteotomic site is distracted for 7days. Consolidation period is 28days. The animal was sacrificed at the 3rd, 7th, 14th, 28th day after the operation. The distracted bone was examined by histological analysis and RT-PCR analysis. The results were summarized as follows: 1. Experimental group was observed that the gaps between the distracted bone edges were occupied by new bone. 2. Expression of Type I collagen were detected throughout the experiment in both groups and Expression of Type I collagen were markedly increased during distraction and consolidation period in experimental group than control group. 3. Expression of Type II collagen were detected throughout the experiment in both groups and expression of Type II collagen were maintained at high level during distraction and consolidation period in experimental group than control group. From these results, in contrast to type II collagen, type I collagen seemed to be more expressed by mechanical stimuli during distraction and consolidation period. The predominent mechanism of new bone formation in the distraction gap was intramembranous bone formation, but some of the regenerated bone was formed by endochondral ossification.
Zeng, Wen-Li;Chen, Yao-Wu;Zhou, Hui;Zhou, Jue-Yu;Wei, Min;Shi, Rong
Asian Pacific Journal of Cancer Prevention
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제16권2호
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pp.513-517
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2015
Background: Growing evidence suggests that the members of the ubiquitin-proteasome system (UPS) are important for tumorigenesis. HERC4, one component, is a recently identified ubiqutin ligase. However, the expression level and function role of HERC4 in lung cancer remain unknown. Our objective was to investigate any correlation between HERC4 and development of lung cancer and its clinical significance. Materials and Methods: To determine HERC4 expression in lung cancer, an immunohistochemistry analysis of a tissue microarray containing samples of 10 lung normal tissues, 15 pulmonary neuroendocrine carcinomas, 45 squamous epithelial cancers and 50 adenocarcinomas was conducted. Receiver operating characteristic (ROC) curve analysis was applied to obtain a cut-off point of 52.5%, above which the expression of HERC4 was regarded as "positive". Results: On the basis of ROC curve analysis, positive expression of HERC4 was detected in 0/10 (0.0%) of lung normal tissues, in 4/15 (26.7%) of pulmonary neuroendocrine carcinomas, in 13/45 (28.9%) of squamous epithelial cancers and in 19/50 (38.0%) of adenocarcinomas. It showed that lung tumors expressed more HERC4 protein than adjacent normal tissues (${\chi}^2$=4.675, p=0.031). Furthermore, HERC4 positive expression had positive correlation with pT status (${\chi}^2$=44.894, p=0.000), pN status (${\chi}^2$=43.628, p=0.000), histological grade (${\chi}^2$=7.083, p=0.029) and clinical stage (${\chi}^2$=72.484, p=0.000), but not age (${\chi}^2$=0.910, p=0.340). Conclusions: Our analysis suggested that HERC4 is likely to be a diagnostic biomarker for lung cancer.
Ji, Hong;Wang, Jianfa;Liu, Juxiong;Guo, Jingru;Wang, Zhongwei;Zhang, Xu;Guo, Li;Yang, Huanmin
Asian-Australasian Journal of Animal Sciences
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제26권3호
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pp.423-432
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2013
Zi geese (Anser anser domestica) belong to the white geese and are excellent layers with a superior feed-to-egg conversion ratio. Quantitative gene expression analysis, such as Real-time qRT-PCR, will provide a good understanding of ovarian function during egg-laying and consequently improve egg production. However, we still don't know what reference genes in geese, which show stable expression, should be used for such quantitative analysis. In order to reveal such reference genes, the stability of seven genes were tested in five tissues of Zi geese. Methodology/Principal Findings: The relative transcription levels of genes encoding hypoxanthine guanine phosphoribosyl transferase 1 (HPRT1), ${\beta}$-actin (ACTB), ${\beta}$-tubulin (TUB), glyceraldehyde-3-phosphate-dehydrogenase (GADPH), succinate dehydrogenase flavoprotein (SDH), 28S rRNA (28S) and 18S rRNA (18S) have been quantified in heart, liver, kidney, muscle and ovary in Zi geese respectively at different developmental stages (1 d, 2, 4, 6 and 8 months). The expression stability of these genes was analyzed using geNorm, NormFinder and BestKeeper software. Conclusions: The expression of 28S in heart, GAPDH in liver and ovary, ACTB in kidney and HPRT1 in muscle are the most stable genes as identified by the three different analysis methods. Thus, these genes are recommended for use as candidate reference genes to compare mRNA transcription in various developmental stages of geese.
2-Oxoglutarate (2OG) acts as a signaling molecule and plays a critical role in secondary metabolism in a variety of organisms, including plants. Six 2-oxoglutarate (2OG) and Fe(II) oxygenase (2OGO) genes, VlCE2OGO1 [Vitis labruscana 2-oxoglutarate (2OG) and Fe(II) oxygenase 1], VlCE2OGO2, VlCE2OGO3, VlCE2OGO4, VlCE2OGO5, and VlCE2OGO6, which show different expression patterns upon transcriptome analysis of 'Campbell Early' grapevine exposed to low temperature for 4 weeks, were analyzed for their structure and expression. Comparison of the deduced amino acid sequences of the 2OGO genes from the V. labruscana transcripts revealed sequence similarities of 38.6% (VlCE2OGO1 and VlCE2OGO2) to 19.2% (VlCE2OGO2 and VlCE2OGO3). The lengths of these genes ranged from 1053 to 2298 bp, and they encoded 316 to 380 amino acids. The prediction of the secondary structure of the encoded proteins by Self-Optimized Prediction Method with Alignment (SOPMA) indicated that all the genes contained alpha helix (23.95 to 41.71%), extended strand (16 to 22.34%), beta turn (6.65 to 9.22%), and random coil (32.97 to 51.58%) in the analysis. Specific primers from unique regions in each gene obtained by alignment of nucleotide sequences were used in real time PCR for analysis of gene expression. All tested genes showed differential expression in grapevines exposed to low temperature. Of the six transcripts, VlCE2OGO1, VlCE2OGO2, and VlCE2OGO3 were up-regulated and VlCE2OGO4, VlCE2OGO5, and VlCE2OGO6 were down-regulated in response to cold treatments at all tested time points. The 2OG genes can be used for elucidation of mechanisms of tolerance to cold and as valuable molecular genetic resources for selection in breeding programs for cold-hardy grapevines.
Won-Sik Woo;Kwang-Min Choi;Min-Soo Joo;Gyoungsik Kang;Kyung-Ho Kim;Ha-Jeong Son;Min-Young Sohn;Do-Hyung Kim;Chan-Il Park
Fisheries and Aquatic Sciences
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제26권2호
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pp.97-104
/
2023
CD96 is a membrane-bound receptor discovered in humans in 1992 that is mainly present in natural killer cells and T cells derived from haematopoietic cells and performs immune functions. Based on the sequence of CD96 obtained from red seabream (Pagrus major), phylogenetic analysis with other species, infections of normal fish, Streptococcus iniae and red sea bream iridovirus (RSIV), and expression analysis was conducted using real-time polymerase chain reaction. Phylogenetic analysis showed the highest homology with Sparus aurata, and multiple sequence analysis confirmed the conservation of major domains between different fish species. Normal fish high expression results were confirmed in the head kidney, and spleen, which are the haematopoietic organs of the fish. High expression levels were confirmed in the gills, liver, spleen, and kidney on day three after RSIV infection. After S. iniae infection, high expression was confirmed in the gills and liver on day one, and high expression was confirmed in the spleen from 12 hours. These results show that PmCD96 functions as an immune gene in P. major and is considered a basic research case for CD96 in fish's hematopoietic organ immune system.
Lee, Young Mee;Lee, Jeong-Ho;Noh, Jae Koo;Kim, Hyun Chul;Park, Choul-Ji;Park, Jong-Won;Hwang, In Joon;Kim, Sung Yeon
한국발생생물학회지:발생과생식
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제17권4호
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pp.329-335
/
2013
TCR subunits are members of membrane-bound receptors which allow the fast and efficient elimination of the specific fish pathogens have regulated function in adaptive immunity. Sequence structure of TCR subunits have been reported for various teleosts, but the information of each TCR subunit functional characterization through expression analysis in fish was unknown. In this study, we examined the gene expression of TCR subunits in the early developmental stages and observed transcript levels in various tissues from healthy adult olive flounder by RT-PCR. The mRNA expression of alpha subunit was already detected in the previous hatching step. But the transcripts of another TCR subunit were not observed during embryo development and increased after hatching and maintained until metamorphosis at the same level. It was found that all TCR subunits mRNAs are commonly expressed in the immune-related organ such as spleen, kidney and gill, also weak expressed in fin and eye. TCR alpha and beta subunit were expressed in brain, whereas gamma and delta were not expressed same tissue. The sequence alignment analysis shows that there are more than 80% sequence homology between TCR subunits. Because it has a high similarity of amino acid sequence to expect similar in function, but expression analysis show that will have may functional diversity due to different time and place of expression.
Background: MicroRNAs are a class of noncoding RNAs which regulate multiple cellular processes during tumor development. The purpose of this report is to investigate the clinicopathological and prognostic significance of miR-218 in human gliomas. Materials and Methods: Quantitative RT-PCR (qRT-PCR) was conducted to detect the expression of miR-218 in primary normal human astrocytes, three glioma cell lines and 98 paired glioma and adjacent normal brain tissues.Associations of miR-218 with clinicopathological variables of glioma patients were statistically analyzed. Finally, a survival analysis was performed using the Kaplan-Meier method and Cox's proportional hazards model. Results: The expression level of miR-218 in primary normal human astrocytes was significantly higher than that in glioma cell lines (p<0.01). Also, the expression level of miR-218 in glioma tissues was significantly downregulated in comparison with that in the adjacent normal brain tissues (p<0.001). Statistical analyses demonstrated that low miR-218 expression was closely associated with advanced WHO grade (p=0.002) and low Karnofsky performance score (p=0.010) of glioma patients. Kaplan-Meier analysis with the log-rank test showed that patients with low-miR-218 expression had poorer disease-free survival and overall survival (p=0.0045 and 0.0124, respectively). Multivariate analysis revealed that miR-218 expression was independently associated with the disease-free survival (p=0.009) and overall survival (p=0.004) of glioma patients. Conclusions: Our results indicate that miR-218 is downregulated in gliomas and that its status might be a potential valuable biomarker for glioma patients.
Abel et al. in Germany discovered a new dioxin-responsive gene, which has later been identified as rpt-1 (regulatory protein T-lymphocyte 1). While it is speculated that rpt-1 may play a role in signal transduction and carcinogenesis, its roles and functions remain unknown. The present study attempted to analyze functions of rpt-1 in human epithelial cells following the xenobiotic exposures. While German counterpart analyzed expressionn of rpt-1 in spleen and thymus cells from mouse and rat and characterizes molecular properties of the gene, our work mainly focused on analyzing function of rpt-1 in human skin cells. Expression of rpt-1 in human cells were analyzed by western and northern blot RT-PCR analysis. Expression of rpt-1 as well as Staf-50 in human cells with or without exposure to environmental pollutants were also analyzed by northern blot analysis, since Staf-50 is homologous with rpt-1 and found in human cells. To help study roles of rpt-1 in human cell system, retroviral vector system carrying rpt-1 gene under the CMV promoter were constructed and transfected. Cells overexpressing the gene after the transfection showed an increase of cell density and soft agar colony formations, as compared to the control cells, suggesting that rpt-1 may play a certain role in the transformation processes of human cells. While the expression of rpt-1 in spleen and thymus is known to be strong in the laboratory animals, both the basal and TCDD-induced expression of rpt-1 in the current cellular system remained insignificant. It is speculated that the expression pattern of rpt-1 may be tissue- and species-specific. The present study demonstrated a strong expression of rpt-1 protein in the brain of SD rat model. Since there is no previous report on the expression of rpt-1 in the brain tissue, the result may play a significant role in understanding dioxin-induced neurotoxicities in the future. The present study provides an opportunity to understand a role of rpt-1 in human cell system and suggest a possible lead and basis for the future study of dioxin-induced neurotoxicities.
Purpose: Dysregulation of apoptosis may attribute to development of cancer by abnormally prolonging cell viability with accumulation of transforming mutations. Survivin and HIAP (Human Inhibitors of Apoptosis)-1 were recently described as apoptosis inhibitors. Their pathogenic roles in gastric cancer are largely unknown. In the present study, we examined the expression of survivin and HIAP-1 in gastric cancer tissues and cell lines in order to elucidate the roles of survivin and HIAP-1 in the process of gastric carcinogenesis. Materials and Methods: Eight gastric cancer cell lines and five gastric cancer tissues were studied. The expression of survivin and HIAP-1 were evaluated by reverse transcription -polymerase chain reaction (RT-PCR), immunohistochemistry, and Western blot. Results: Western blot and RT-PCR analysis revealed survivin and HIAP-1 expression in all gastric cancer cell lines. Increased expression of survivin and HIAP-1 were found in all cases of gastric cancer tissues compared to normal tissues by Western blot analysis. In immunohistochemical analysis tumor cells were stained with anti-survivin and anti-HIAP-1 antibodies. Cell cycle dependence of survivin expression was preserved in gastric cancer cell lines. Conclusion: The results indicate that increased expression of survivin and HIAP-1 genes may play an important role in gastric cancer.
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