Multiple signal transduction pathways regulate clusterin (gp 80) gene expression in MDCK cells

Gutacker, Claudia; Flach, Rachel J. Roth; Diel, Patrick; Klöck, Gerd; Koch-Brandt, Claudia

doi:10.1677/jme.0.0170109

Cited by 13 publications

(14 citation statements)

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“…The findings of clusterin induction by EGF in PC12 cells contrasts with previous observations that EGF repressed clusterin mRNA in the kidney-derived cell line Madin-Darby canine kidney (MDCK) [26]. Similarly, EGF was shown to down-regulate clusterin expression in the polycystic kidneys of mice [36].…”

Section: Discussioncontrasting

confidence: 95%

“…Moreover, clusterin expression is associated with differentiation and development [35] : the gene is activated in the mouse embryo at sites of epithelial differentiation [6] and responds to proliferative stimuli in a variety of systems [26,[36][37][38]. Only little information is available, however, on the molecular details of clusterin gene regulation in these processes.…”

Section: Discussionmentioning

confidence: 99%

“…NGF and EGF, both from mouse submaxillary gland, were obtained from Boehringer Mannheim and added to the culture medium as indicated. Total RNA isolation and Northern blotting was done as described [26]. Each sample of total RNA (10 µg) was run on 1.2 % (w\v) agarose\0.66 M formaldehyde gels and blotted to nylon membranes (Boehringer, Mannheim).…”

Section: Experimental Cell Culture Rna Isolation and Northern Blottingmentioning

confidence: 99%

“…At day 0 the medium was changed to serum-containing medium plus 100 ng/ml NGF. At the indicated time points the cells were harvested and RNA was isolated as described [26]. A Northern blot was performed with 10 µg of total RNA in each lane ; the blot was hybridized to a radioactively labelled probe of the rat clusterin gene (SGP-2 [26]) ; after autoradiography, the clusterin probe was removed and the blot was rehybridized with a probe for the β-actin gene.…”

Section: Figure 2 Ngf Activates Clusterin Mrna In Pc12 Cellsmentioning

confidence: 99%

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Nerve growth factor and epidermal growth factor stimulate clusterin gene expression in PC12 cells

Gutacker

Klöck

DIEL³

et al. 1999

Biochemical Journal

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Clusterin (apolipoprotein J) is an extracellular glycoprotein that might exert functions in development, cell death and lipid transport. Clusterin gene expression is elevated at sites of tissue remodelling, such as differentiation and apoptosis; however, the signals responsible for this regulation have not been identified. We use here the clusterin gene as a model system to examine expression in PC12 cells under the control of differentiation and proliferation signals produced by nerve growth factor (NGF) and by epidermal growth factor (EGF) respectively. NGF induced clusterin mRNA, which preceded neurite outgrowth typical of neuronal differentiation. EGF also activated the clusterin mRNA, demonstrating that both proliferation and differentiation signals regulate the gene. To localize NGF- and EGF-responsive elements we isolated the clusterin promoter and tested it in PC12 cell transfections. A 2.5 kb promoter fragment and two 1.5 and 0.3 kb deletion mutants were inducible by NGF and EGF. The contribution to this response of a conserved activator protein 1 (AP-1) motif located in the 0.3 kb fragment was analysed by mutagenesis. The mutant promoter was not inducible by NGF or EGF, which identifies the AP-1 motif as an element responding to both factors. Binding studies with PC12 nuclear extracts showed that AP-1 binds to this sequence in the clusterin promoter. These findings suggest that NGF and EGF, which give differential gene regulation in PC12 cells, resulting in neuronal differentiation and proliferation respectively, use the common Ras/extracellular signal-regulated kinase/AP-1 signalling pathway to activate clusterin expression.

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Section: Discussioncontrasting

confidence: 95%

Section: Discussionmentioning

confidence: 99%

Section: Experimental Cell Culture Rna Isolation and Northern Blottingmentioning

confidence: 99%

Section: Figure 2 Ngf Activates Clusterin Mrna In Pc12 Cellsmentioning

confidence: 99%

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Nerve growth factor and epidermal growth factor stimulate clusterin gene expression in PC12 cells

Gutacker

Klöck

DIEL³

et al. 1999

Biochemical Journal

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“…Clusterin expression is known to be suppressed by gonadal steroids in prostate, mammary gland and uterus (Bettuzzi et al 1992, Wunsche et al 1998 and by glucocorticoids in brain and kidney cells (Finch & May 1995, Gutacker et al 1996. These observations led us to investigate clusterin expression in the ovine pituitary gland, with the aim of determining the distribution of clusterin mRNA and protein in the different parts of this tissue.…”

Section: Introductionmentioning

confidence: 99%

Clusterin is expressed in the anterior and intermediate lobes, but not in the posterior pituitary of sheep

Fleming¹,

Hope²,

Bolter³

1999

Journal of Molecular Endocrinology

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We have examined the expression of the ovine clusterin gene in the sheep pituitary gland, with the aim of determining its site of synthesis in this tissue. Northern blotting analysis of extracted polyadenylated RNA, using a 32 P-labelled rat clusterin cDNA probe, detected the greatest amounts of clusterin mRNA in the anterior part of dissected pituitary glands. In situ hybridisation studies showed clusterin mRNA in anterior and intermediate Journal of Molecular Endocrinology (1999) 23, 199-208 INTRODUCTIONClusterin is a highly sulphated glycoprotein that is expressed in a wide range of tissues, often in association with tissue remodelling or apoptosis, in response to injury or pathology, or connected with lipid transport or atherosclerotic vascular disease (Fritz & Murphy 1993). It is expressed in many endocrine tissues, including ovary, testis, prostate and uterus (Sylvester & Griswold 1995), pancreas (Scaglia et al. 1995) and breast tissue (Tenniswood et al. 1992, Welsh 1994. Clusterin is known to be an adrenal chromaffin granule constituent and its expression increases in the adrenal in response to histamine or forskolin treatment (Palmer & Christie 1990, Laslop et al. 1993. In gonadal tissue, large amounts of clusterin mRNA have been measured in extracts of ovine corpora lutea (Fleming et al. 1992) and high concentrations of clusterin protein are synthesised in the ram rete testis (Tung & Fritz 1985).There is further evidence for a role for this protein in search of a function (Fritz 1995) in neural tissue, where its expression is upregulated in pathologies such as Alzheimer's disease (Finch & May 1995). Clusterin is induced in reactive astrocytes in response to neurotoxic injury (Zwain et al. 1994, Walton et al. 1996, Tornquist et al. 1997 and appears to have a protective role in the brain, possibly acting as a local inhibitor of the complement cascade (Tornquist et al. 1996).Clusterin expression is known to be suppressed by gonadal steroids in prostate, mammary gland and uterus (Bettuzzi et al. 1992, Wunsche et al. 1998 and by glucocorticoids in brain and kidney cells (Finch & May 1995, Gutacker et al. 1996. These observations led us to investigate clusterin expression in the ovine pituitary gland, with the aim of determining the distribution of clusterin mRNA and protein in the different parts of this tissue. In previous studies of bovine pituitary, the protein was detected in both the anterior and posterior lobes (Laslop et al. 1993). However, the site of clusterin mRNA transcription remains unclear. Aronow et al. (1993) did not detect clusterin mRNA in mouse anterior pituitary, whereas Laslop et al. (1993) (Fleming et al. 1997) was used as a positive control on Northern blots probed with the growth hormone gene probe.Pituitary glands for immunohistochemistry and in situ hybridisation were obtained from five 4-to 6-year-old Romney ewes killed with Euthatal (Southern Veterinary Supplies, Christchurch, New Zealand; 100 mg/kg, i.v.), at the mid-luteal phase of the oestrous cycle. Whole glands were ...

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Differential regulation of the clusterin gene by Ha-ras and c-myc oncogenes and during apoptosis

Klöck

Storch

Rickert³

et al. 1998

J. Cell. Phys.

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Clusterin (ApoJ) is an extracellular glycoprotein expressed during processes of tissue differentiation and regression that involve programmed cell death (apoptosis). Increased clusterin expression has also been found in tumors, however, the mechanism underlying this induction is not known. Apoptotic processes in tumors could be responsible for clusterin gene activation. Alternatively, oncogenic mutations could modulate signal transduction, thereby inducing the gene. We examined the response of the rat clusterin gene to two oncogenes, Ha-ras and c-myc, in transfected Rat1 fibroblasts. While c-myc overexpression did not modify clusterin gene activity, the Ha-ras oncogene produced a seven to tenfold repression of clusterin mRNA; this down-regulation was also observed in the presence of c-myc. Since no induction of the clusterin gene was observed by the two oncogenes, we tested the alternative mechanism involving apoptosis. Growth factor withdrawal induced apoptosis, as shown by DNA degradation and micronuclei formation in the floating cells. Concomittantly we observed a three to tenfold increase in the amount of clusterin mRNA in the adhering cells of Rat1 and the c-myc transformed cell lines, and a weaker induction in the Ha-ras transformed cell line. On the basis of our results, we suggest that clusterin gene induction in the vital cells is produced by signaling molecules that are generated by the apoptotic cells. We conclude that apoptotic processes, not oncogenic mutations, are responsible for increased clusterin expression in tumors.

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Multiple signal transduction pathways regulate clusterin (gp 80) gene expression in MDCK cells

Cited by 13 publications

References 0 publications

Nerve growth factor and epidermal growth factor stimulate clusterin gene expression in PC12 cells

Nerve growth factor and epidermal growth factor stimulate clusterin gene expression in PC12 cells

Clusterin is expressed in the anterior and intermediate lobes, but not in the posterior pituitary of sheep

Differential regulation of the clusterin gene by Ha-ras and c-myc oncogenes and during apoptosis

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