• Title/Summary/Keyword: quantitative real-time RT-PCR

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Effects of Coenzyme Q10 on the Expression of Genes involved in Lipid Metabolism in Laying Hens (Coenzyme Q10 첨가 급여가 산란계의 지방대사 연관 유전자 발현에 미치는 영향)

  • Jang, In Surk;Moon, Yang Soo
    • Korean Journal of Poultry Science
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    • v.43 no.1
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    • pp.47-54
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    • 2016
  • The aim of this study was to investigate the expression patterns of key genes involved in lipid metabolism in response to dietary Coenzyme Q10 (CoQ10) in hens. A total of 36 forty week-old Lohmann Brown were randomly allocated into 3 groups consisting of 4 replicates of 3 birds. Laying hens were subjected to one of following treatments: Control (BD, basal diet), T1 (BD+ CoQ10 100 mg/kg diet) and T2 (BD+ micellar of CoQ10 100 mg/kg diet). Birds were fed ad libitum a basal diet or the basal diet supplemented with CoQ10 for 5 weeks. Total RNA was extracted from the liver for quantitative RT-PCR. The mRNA levels of HMG-CoA reductase(HMGCR) and sterol regulatory element-binding proteins(SREBP)2 were decreased more than 30~50% in the liver of birds fed a basal diet supplemented with CoQ10 (p<0.05). These findings suggest that dietary CoQ10 can reduce cholesterol levels by the suppression of the hepatic HMGCR and SREBP2 genes. The gene expressions of liver X receptor (LXR) and SREBP1 were down regulated due to the addition of CoQ10 to the feed (p<0.05). The homeostasis of cholesterol can be regulated by LXR and SREBP1 in cholesterol-low-conditions. The supplement of CoQ10 caused a decreased expression of lipid metabolism-related genes including $PPAR{\gamma}$, XBP1, FASN, and GLUTs in the liver of birds (p<0.05). These data suggest that CoQ10 might be used as a dietary supplement to reduce cholesterol levels and to regulate lipid homeostasis in laying hens.

Microarray Analysis of Long Non-coding RNA Expression Profile Associated with 5-Fluorouracil-Based Chemoradiation Resistance in Colorectal Cancer Cells

  • Xiong, Wei;Jiang, Yong-Xin;Ai, Yi-Qin;Liu, Shan;Wu, Xing-Rao;Cui, Jian-Guo;Qin, Ji-Yong;Liu, Yan;Xia, Yao-Xiong;Ju, Yun-He;He, Wen-Jie;Wang, Yong;Li, Yun-Fen;Hou, Yu;Wang, Li;Li, Wen-Hui
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.8
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    • pp.3395-3402
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    • 2015
  • Background: Preoperative 5-fluorouracil (5-FU)-based chemoradiotherapy is a standard treatment for locally advanced colorectal cancer (CRC). However, CRC cells often develop chemoradiation resistance (CRR). Recent studies have shown that long non-coding RNA (lncRNA) plays critical roles in a myriad of biological processes and human diseases, as well as chemotherapy resistance. Since the roles of lncRNAs in 5-FU-based CRR in human CRC cells remain unknown, they were investigated in this study. Materials and Methods: A 5-FU-based concurrent CRR cell model was established using human CRC cell line HCT116. Microarray expression profiling of lncRNAs and mRNAs was undertaken in parental HCT116 and 5-FU-based CRR cell lines. Results: In total, 2,662 differentially expressed lncRNAs and 2,398 mRNAs were identified in 5-FU-based CRR HCT116 cells when compared with those in parental HCT116. Moreover, 6 lncRNAs and 6 mRNAs found to be differentially expressed were validated by quantitative real time PCR (qRT-PCR). Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis for the differentially expressed mRNAs indicated involvement of many, such as Jak-STAT, PI3K-Akt and NF-kappa B signaling pathways. To better understand the molecular basis of 5-FU-based CRR in CRC cells, correlated expression networks were constructed based on 8 intergenic lncRNAs and their nearby coding genes. Conclusions: Changes in lncRNA expression are involved in 5-FU-based CRR in CRC cells. These findings may provide novel insight for the prognosis and prediction of response to therapy in CRC patients.

Effects of Lycopene on the Expression of Lipid Metabolism, Glucose Transport and Pro-Inflammatory Related Genes in Chickens (라이코펜 첨가 급여가 닭의 지방대사, 포도당 수송 및 친염증 유전자 발현에 미치는 영향)

  • Jang, In Surk;Moon, Yang Soo
    • Korean Journal of Poultry Science
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    • v.42 no.3
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    • pp.231-238
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    • 2015
  • The present study aimed to investigate the effects of lycopene on hepatic metabolic- and immune-related gene expression in laying hens. A total of 48 25-week-old White Leghorn hens were randomly allocated into four groups consisting of four replicates of three birds: control (basal diet), T1 (basal diet + 10 mg/kg of tomato powder-containing lycopene), T2 (basal diet + 10 mg/kg of micelles of tomato powder-containing lycopene), and T3 (basal diet + 10 mg/kg of purified lycopene). Chickens were fed ad libitum for 5 weeks, and then total RNA was extracted from the livers for quantitative RT-PCR analysis. Peroxisome proliferator-activated receptor ${\gamma}$ (PPAR${\gamma}$) expression was decreased in the liver of chickens after lycopene supplementation (P<0.05). Micellar lycopene supplementation decreased the expression of PPAR${\gamma}$ target genes including fatty acid binding protein 4 (FABP4) and fatty acids synthase (FASN) in the T2 group (P<0.05). Sterol regulatory element-binding protein 2 (SREBP2) and C/EBP-${\alpha}$ were also downregulated in hens fed with micellar lycopene (P<0.05). Glucose transporter 8 (GLUT-8) was upregulated in the T2 and T3 groups (P<0.05). However, the expression of carnitine palmitoyltransferase 1 (CPT-1) was not changed by lycopene supplementation. Pro-inflammatory cytokines such as tumor necrosis factor ${\alpha}$ (TNF-${\alpha}$) and interleukin 6 (IL-6) were downregulated by lycopene supplementation (P<0.05). These data suggest that the type of lycopene supplementation is critical and that micelles of tomato powder-containing lycopene may play an important role in the modulation of lipid metabolism and immunity in chickens.

Expression of Osteopontin in Eutopic and Ectopic Endometrial Tissues in Endometriosis (자궁내막증 환자의 정상위치 및 이소성 자궁내막에서의 Osteopontin의 발현)

  • Koo, Yun-Hee;Kim, Chung-Hoon;Kim, Ji-Sun;Lee, Young-Jin;Kim, Sung-Hoon;Chae, Hee-Dong;Kang, Byung-Moon
    • Clinical and Experimental Reproductive Medicine
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    • v.34 no.3
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    • pp.149-157
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    • 2007
  • Objective: This study was performed to compare the expression of osteopontin (OPN) mRNA and protein in the eutopic and ectopic endometrial tissues in women with endometriosis and endometrial tissues in women without endometriosis. Methods: A total of 32 women with histologically confirmed endometriosis were recruited for study group. For controls, 34 women undergoing operative treatment for cervical intraepithelial neoplasia (CIN) or benign gynecologic condition other than endometriosis were recruited. At the time of laparoscopy or laparotomy, a biopsy specimen was taken from the endometrial cavity and peritoneal endometrial implant or endometrioma whenever appropriate. We employed real time quantitative RT-PCR to quantify OPN mRNA expression of these tissues and performed western blot analysis to measure the quantity of OPN. Results: The expression of OPN mRNA was significantly higher in both eutopic and ectopic endometrial tissues of women with endometriosis than in endometrial tissues of controls during both proliferative and secretory phase. In the eutopic endometrial tissue of women with endometriosis, OPN mRNA expression significantly increased during the secretory phase compared to the proliferative phase in women with endometriosis as well as controls. However, in the ectopic endometrial tissue, OPN mRNA expression significantly decreased during the secretory phase compared to the proliferative phase. The expression of OPN protein was significantly higher in women with endometriosis than in controls. Conclusion: This study shows the marked expression of OPN mRNA and protein in eutopic and ectopic endometrial tissues in women with endometriosis may be associated with the adhesion and invasion of endometrial explants.

Gene Expression Profile Associated with the Differentiation of Osteoblasts from Human Mesenchymal Stem Cells (인간 중간엽 줄기세포로부터 골아세포로의 분화시 관찰되는 유전자 발현 분석)

  • Kim Yeo-Kyeoung;Kim Hee-Nam;Lee Il-Kwon;Park Kyeong-Soo;Yang Deok-Hwan;Cho Sang-Hee;Lee Je-Jung;Chung Ik-Joo;Kim Soon-Hag;Kim Hyeoung-Joon
    • Journal of Life Science
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    • v.16 no.2 s.75
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    • pp.231-239
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    • 2006
  • Human mesenchymal stem cells (hMSCs) in bone marrow (BM) can be induced to differentiate into a variety of mesenchymal tissues, including adipocytes, osteoblasts and chondroblasts, under the influence of certain growth or environmental factors. In this study, we analyzed the differentiation process and the associated gene expression profiles inherent to the process by which hMSCs differentiate into osteoblasts. We conducted a comparison of gene expression profiles of the normal human BM MSCs, using human 8K cDNA microarray, incubated in media containing either a combination of $\beta$-glycerol phosphate, L-ascorbic acid, and dexamethasone, or in medium lacking these osteogenic supplements. During the osteoblastic differentiation process, 36 genes were determined to be up-regulated, and 59 genes were shown to be down-regulated. Osteoprotegerin, LRP5, and metallothionein 2A, all of which are associated with the osteogenetic process, were up-regulated, and genes associated with the differentiation of MSCs into other lineages, including muscle, adipose tissue and vascular structure were down-regulated. The set of differentially expressed genes reported in this work should contribute to our current understanding of the processes inherent to the differentiation of MSCs into osteoblasts.