Volume 127, Issue 4, Pages (October 2004)

Slides:



Advertisements
Similar presentations
IL-18 Downregulates Collagen Production in Human Dermal Fibroblasts via the ERK Pathway  Hee Jung Kim, Seok Bean Song, Jung Min Choi, Kyung Moon Kim,
Advertisements

Volume 131, Issue 4, Pages (October 2006)
MicroRNA-558 regulates the expression of cyclooxygenase-2 and IL-1β-induced catabolic effects in human articular chondrocytes  S.J. Park, E.J. Cheon,
Volume 144, Issue 3, Pages e4 (March 2013)
Mira Park, Ph. D. , Dae-Shik Suh, M. D. , Kangseok Lee, Ph. D
Volume 35, Issue 4, Pages (August 2009)
Zhao Jing, M. D. , Zhang Qiong, M. D. , Wang Yonggang, M. D
Microsomal Prostaglandin E Synthase-1 Inhibits PTEN and Promotes Experimental Cholangiocarcinogenesis and Tumor Progression  Dongdong Lu, Chang Han, Tong.
Volume 134, Issue 4, Pages (April 2008)
Takashi Tanaka, Michelle A. Soriano, Michael J. Grusby  Immunity 
Volume 137, Issue 2, Pages e2 (August 2009)
IL-2–mediated apoptosis of kidney tubular epithelial cells is regulated by the caspase-8 inhibitor c-FLIP  Caigan Du, Qiunong Guan, Ziqin Yin, Robert.
Volume 136, Issue 3, Pages e2 (March 2009)
Richard T. Ethridge, Mark R. Hellmich, Raymond N. DuBois, B.Mark Evers 
Volume 126, Issue 3, Pages (March 2004)
Volume 133, Issue 6, Pages (December 2007)
MicroRNA-558 regulates the expression of cyclooxygenase-2 and IL-1β-induced catabolic effects in human articular chondrocytes  S.J. Park, E.J. Cheon,
Volume 150, Issue 1, Pages e4 (January 2016)
Teruaki Fujishita, Masahiro Aoki, Makoto M. Taketo  Gastroenterology 
Volume 115, Issue 6, Pages (December 1998)
Volume 25, Issue 4, Pages (April 2014)
Volume 3, Issue 1, Pages (January 1999)
Volume 132, Issue 3, Pages (March 2007)
Zinc Mesoporphyrin Induces Rapid Proteasomal Degradation of Hepatitis C Nonstructural 5A Protein in Human Hepatoma Cells  Weihong Hou, Qing Tian, Jianyu.
David X Liu, Lloyd A Greene  Neuron 
Volume 62, Issue 4, Pages (October 2002)
Volume 141, Issue 5, Pages (November 2011)
Volume 131, Issue 4, Pages (October 2006)
IFN-γ Upregulates Expression of the Mouse Complement C1rA Gene in Keratinocytes via IFN-Regulatory Factor-1  Sung June Byun, Ik-Soo Jeon, Hyangkyu Lee,
Volume 137, Issue 3, Pages (September 2009)
Laminin γ2 Mediates Wnt5a-Induced Invasion of Gastric Cancer Cells
Volume 138, Issue 2, Pages e2 (February 2010)
Volume 133, Issue 5, Pages (November 2007)
Volume 137, Issue 4, Pages (October 2009)
Volume 116, Issue 6, Pages (June 1999)
Volume 138, Issue 5, Pages e2 (May 2010)
Volume 141, Issue 4, Pages (October 2011)
Volume 134, Issue 1, Pages (January 2008)
Volume 139, Issue 6, Pages (December 2010)
Volume 130, Issue 1, Pages (January 2006)
Volume 137, Issue 1, Pages (July 2009)
Volume 135, Issue 3, Pages e3 (September 2008)
Mechanisms of cross hyporesponsiveness to toll-like receptor bacterial ligands in intestinal epithelial cells  Jan-Michel Otte, Elke Cario, Daniel K.
Volume 132, Issue 5, Pages (May 2007)
Volume 57, Issue 5, Pages (May 2000)
Histamine Enhances the Production of Granulocyte-Macrophage Colony-Stimulating Factor via Protein Kinase Cα and Extracellular Signal-Regulated Kinase.
Antigen-Presenting Cell Production of IL-10 Inhibits T-Helper 1 and 17 Cell Responses and Suppresses Colitis in Mice  Bo Liu, Susan L. Tonkonogy, R. Balfour.
Microsomal Prostaglandin E Synthase-1 Inhibits PTEN and Promotes Experimental Cholangiocarcinogenesis and Tumor Progression  Dongdong Lu, Chang Han, Tong.
Molecular Therapy - Nucleic Acids
Volume 137, Issue 4, Pages (October 2009)
Ling Zheng, Terrence E. Riehl, William F. Stenson  Gastroenterology 
All-Trans-Retinoic Acid Induces Interleukin-8 via the Nuclear Factor-κB and p38 Mitogen-Activated Protein Kinase Pathways in Normal Human Keratinocytes 
Keratinocyte growth factor promotes goblet cell differentiation through regulation of goblet cell silencer inhibitor  Dai Iwakiri, Daniel K. Podolsky 
17β-estradiol Inhibits the Production of RANTES in Human Keratinocytes
Volume 141, Issue 3, Pages (September 2011)
Volume 128, Issue 3, Pages (March 2005)
Volume 61, Issue 6, Pages (June 2002)
Amphiregulin: An early trigger of liver regeneration in mice
Volume 2, Issue 6, Pages (June 2008)
Volume 127, Issue 4, Pages (October 2004)
Volume 115, Issue 2, Pages (October 2003)
IL-18 Downregulates Collagen Production in Human Dermal Fibroblasts via the ERK Pathway  Hee Jung Kim, Seok Bean Song, Jung Min Choi, Kyung Moon Kim,
Fan Yang, Huafeng Zhang, Yide Mei, Mian Wu  Molecular Cell 
Volume 18, Issue 3, Pages (March 2010)
Volume 136, Issue 7, Pages (June 2009)
Volume 55, Issue 2, Pages (February 1999)
IL-1β induces VEGF, independently of PGE2 induction, mainly through the PI3-K/mTOR pathway in renal mesangial cells  D. Solà-Villà, M. Camacho, R. Solà,
Volume 72, Issue 2, Pages (July 2007)
Volume 118, Issue 6, Pages (June 2000)
Volume 3, Issue 1, Pages (January 1999)
Presentation transcript:

Volume 127, Issue 4, Pages 1139-1149 (October 2004) Apobec-1 protects intestine from radiation injury through posttranscriptional regulation of cyclooxygenase-2 expression  Shrikant Anant, Nabendu Murmu, Courtney W. Houchen, Debnath Mukhopadhyay, Terrence E. Riehl, Stephen G. Young, Aubrey R. Morrison, William F. Stenson, Nicholas O. Davidson  Gastroenterology  Volume 127, Issue 4, Pages 1139-1149 (October 2004) DOI: 10.1053/j.gastro.2004.06.022 Copyright © 2004 American Gastroenterological Association Terms and Conditions

Figure 1 Stem cell survival in APOBEC-1−/− mice is reduced after radiation injury. (A) Experimental protocol. WT, apoB100-only, and APOBEC-1−/− mice were treated with 12 Gy of γ-irradiation; 82 hours after radiation, BrdU was administered, and the mice were killed 2 hours later. (B) The distal jejunum was stained with H&E for morphological analysis. To identify dividing cells, the tissue was immunostained for BrdU and counterstained with eosin. (C) Total number of regenerating crypts per cross section; values are mean ± SEM (n = 5 mice per genotype). (D) Number of BrdU-positive cells. Values are mean ± SEM (n = 5 mice per genotype). (E) Total number of regenerating crypts per cross section/time course. Values are mean ± SEM (n = 4 mice per genotype). (F) Total number of regenerating crypts/dose curve. Values are mean ± SEM (n = 4 mice per genotype). (G) Epithelial cells at positions 3 to 8 from the base of the crypts are preferentially protected from apoptosis in WT mice. Gastroenterology 2004 127, 1139-1149DOI: (10.1053/j.gastro.2004.06.022) Copyright © 2004 American Gastroenterological Association Terms and Conditions

Figure 1 Stem cell survival in APOBEC-1−/− mice is reduced after radiation injury. (A) Experimental protocol. WT, apoB100-only, and APOBEC-1−/− mice were treated with 12 Gy of γ-irradiation; 82 hours after radiation, BrdU was administered, and the mice were killed 2 hours later. (B) The distal jejunum was stained with H&E for morphological analysis. To identify dividing cells, the tissue was immunostained for BrdU and counterstained with eosin. (C) Total number of regenerating crypts per cross section; values are mean ± SEM (n = 5 mice per genotype). (D) Number of BrdU-positive cells. Values are mean ± SEM (n = 5 mice per genotype). (E) Total number of regenerating crypts per cross section/time course. Values are mean ± SEM (n = 4 mice per genotype). (F) Total number of regenerating crypts/dose curve. Values are mean ± SEM (n = 4 mice per genotype). (G) Epithelial cells at positions 3 to 8 from the base of the crypts are preferentially protected from apoptosis in WT mice. Gastroenterology 2004 127, 1139-1149DOI: (10.1053/j.gastro.2004.06.022) Copyright © 2004 American Gastroenterological Association Terms and Conditions

Figure 2 LPSprotects WT but not APOBEC-1−/− mice from radiation-induced apoptosis. (A) Western blot analyses for COX-2 and HSP40 proteins of lysates from the distal jejunum of mice subjected to 12 Gy of γ-irradiation. The mobility of the molecular weight markers is shown on the left. (B) Schematic representation of treatment regimen. Mice were injected with LPS (10 mg/kg) or dimethyl PGE2 (0.5 mg/kg) 14 hours before γ-irradiation (12 Gy). The mice were killed 6 hours later, and distal jejunum was collected for assessment of apoptosis and PGE2 levels. (C) Radiation-induced apoptosis was assessed by counting the total number of apoptotic cells per crypt in WT and APOBEC-1−/− mice. Values are mean ± SEM (n = 5 mice per genotype). (D) Intestinal PGE2, expressed as mean ± SEM (n = 5 mice per genotype); P <0.001. Gastroenterology 2004 127, 1139-1149DOI: (10.1053/j.gastro.2004.06.022) Copyright © 2004 American Gastroenterological Association Terms and Conditions

Figure 3 COX-2 is not induced in APOBEC-1−/− mice. (A) Mice were treated with LPS 14 hours before irradiation and were killed 6 hours later. Mice treated with LPS only were killed at the time of irradiation. (B) Real-time polymerase chain reaction (PCR) was performed with total RNA from the distal jejunum and expressed as the amount of COX-2 mRNA relative to the glyceraldehyde phosphate dehydrogenase control for each genotype. (C) Lysates of distal jejunum were subjected to Western blot analyses for COX-2, apobec-1, and HSP40 proteins. The mobility of the molecular weight markers is shown on the left. (D) Real-time PCR for c-myc mRNA was performed with total RNA from the distal jejunum and quantitated relative to the glyceraldehyde phosphate dehydrogenase control for each genotype. aP = 0.003. (E) Real-time PCR for c-myc mRNA was performed from distal jejunal RNA from mice subjected to 12 Gy of γ-irradiation. aP <0.0001 compared with WT mice at time 0; bP = 0.002 compared with WT mice. Gastroenterology 2004 127, 1139-1149DOI: (10.1053/j.gastro.2004.06.022) Copyright © 2004 American Gastroenterological Association Terms and Conditions

Figure 4 Apobec-1 interacts with AU-rich sequences in the COX-2 3′ UTR. (A) Electrophoretic mobility shift assay was performed with recombinant glutathione S-transferase (GST)/APOBEC-1 and radiolabeled templates. Probes used in the assay were RB (105 nt of rat apoB RNA containing nt 6639–6743), vascular endothelial growth factor (VEGF; 1.9 kilobases [kb] of 3′ UTR of human VEGF mRNA), COX (2.3 kb of 3′ UTR of mouse COX-2 mRNA), and IL-8 (1.3 kb of 3′ UTR of human IL-8 mRNA). Locations of the apobec-1 bound (b) and free (f) probe are indicated to the right. (B) Apobec-1-bound RNA, isolated from intestinal epithelial cells of WT and APOBEC-1−/− mice exposed to 12 Gy of whole-body γ-irradiation, was subjected to reverse-transcription polymerase chain reaction for COX-2 and glyceraldehyde phosphate dehydrogenase (GAPDH) mRNAs. T, total RNA; P, pellet; S, supernatant. (C) Schematic representation of substitutional mutants generated from the first 60 nt of the COX-2 3′ UTR. Three sets of AU-rich sequences (A-I-A-III) are present in this region. Sequences underlined in each mutant were substituted with the complementary sequences. (D) UV crosslink analysis was performed with recombinant GST/APOBEC-1 and radiolabeled WT and mutant COX-2 3′ UTR transcripts. The arrow indicates the location of the crosslinked band. The mobility of the molecular weight markers is shown on the left. Gastroenterology 2004 127, 1139-1149DOI: (10.1053/j.gastro.2004.06.022) Copyright © 2004 American Gastroenterological Association Terms and Conditions

Figure 5 Apobec-1 stabilizes luciferase (Luc) mRNA containing the first 60 nt of the COX-2 3′ UTR. (A) Schematic representation of Luc plasmids containing the various 3′ UTR regions of the COX-2 mRNA. **AREs within the first 60 nt of COX-2 3′ UTR. (B) The Luc plasmids containing the various COX-2 3′ UTRs were co-transfected with either apobec-1-expressing or β-galactosidase (LacZ)-expressing plasmids into HEK293 cells. Actinomycin D (10 μg/mL) was added to the medium 48 hours after transfection, and the cells were incubated for the indicated times. Total RNA was extracted and subjected to Northern blot analysis for Luc and glyceraldehyde phosphate dehydrogenase mRNAs. Shown is a representative figure of 3 such experiments. (C) The half-life of the Luc mRNAs containing various COX-2 3′ UTRs; values are mean ± SEM of 3 transfections. FL, full length. Gastroenterology 2004 127, 1139-1149DOI: (10.1053/j.gastro.2004.06.022) Copyright © 2004 American Gastroenterological Association Terms and Conditions

Figure 6 The p38 MAPK inhibitor SB202190 (SB) does not affect apobec-1-mediated COX-2 mRNA stability. (A) HeLa cells were infected with recombinant adenovirus constructs carrying β-galactosidase or apobec-1 for 16 hours. SB was added to a final concentration of 20 μmol/L, cells were incubated for a further 6 hours, and lysates were subjected to Western blot analyses for apobec-1 and HSP40. The mobility of the molecular weight markers is shown on the left. (B) Total RNA was isolated at various times after the addition of actinomycin D and subjected to real-time polymerase chain reaction analyses for COX-2 mRNA. The data are presented as COX-2 mRNA levels present relative to those observed at the addition of actinomycin D. Gastroenterology 2004 127, 1139-1149DOI: (10.1053/j.gastro.2004.06.022) Copyright © 2004 American Gastroenterological Association Terms and Conditions