Establishment and immunocharacterization of an immortalized pancreatic cell line derived from the H-2Kb-tsA58 transgenic mouse

Blouin, Richard; Grondin, Gilles; Beaudoin, Jude; Arita, Yoshi; Daigle, Nathalie; Talbot, Brian G.; Lebel, Denis; Morisset, Jean

doi:10.1007/s11626-997-0130-2

Cited by 8 publications

(3 citation statements)

References 24 publications

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“…2). As previously observed in other HIP/PAP expressing cell lines [12,27] the immunostaining generated by anti-HIP/PAP Ig was cytoplasmic and mostly present in the juxta nuclear area (Fig. 2Aa).…”

Section: Identification Of the Riia Regulatory Subunit Of Pka As A Pasupporting

confidence: 86%

HIP/PAP, a C‐type lectin overexpressed in hepatocellular carcinoma, binds the RIIα regulatory subunit of cAMP‐dependent protein kinase and alters the cAMP‐dependent protein kinase signalling

Philippe

Sar

Pileire

et al. 2004

European Journal of Biochemistry

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HIP/PAP is a C-type lectin overexpressed in hepatocellular carcinoma (HCC). Pleiotropic biological activities have been ascribed to this protein, but little is known about the function of HIP/PAP in the liver. In this study, therefore, we searched for proteins interacting with HIP/PAP by screening a HCC cDNA expression library. We have identified the RIIa regulatory subunit of cAMP-dependent protein kinase (PKA) as a partner of HIP/PAP. HIP/PAP and RIIa were coimmunoprecipitated in HIP/PAP expressing cells. The biological relevance of the interaction between these proteins was established by demonstrating, using fractionation methods, that they are located in a same subcellular compartment. Indeed, though HIP/PAP is a protein secreted via the Golgi apparatus we showed that a fraction of HIP/PAP escaped the secretory apparatus and was recovered in the cytosol. Basal PKA activity was increased in HIP/PAP expressing cells, suggesting that HIP/PAP may alter PKA signalling. Indeed, we showed, using a thymidine kinase-luciferase reporter plasmid in which a cAMP responsive element was inserted upstream of the thymidine kinase promoter, that luciferase activity was enhanced in HIP/PAP expressing cells. Thus our findings suggest a novel mechanism for the biological activity of the HIP/PAP lectin.Keywords: C-type lectin; HIP/PAP; PKA; phosphorylation; liver.The HIP/PAP-encoding gene has been shown to be overexpressed in human hepatocellular carcinoma (HCC) [1] and in the pancreas during acute pancreatitis [2]. HIP/ PAP has been characterized as a protein belonging to the group 7 of C-type lectins [3,4]. HIP/PAP cDNA encodes a 175 amino acid protein containing only one carbohydratebinding domain (CRD) linked to an N-terminal sequence, part of which is cleaved during its maturation and secretion [5]. In humans, HIP/PAP protein is not expressed in normal liver but is overexpressed in 75% of HCC, in cholangiocarcinoma and in reactive ductular cells in nonmalignant liver [6]. HIP/PAP expression in HCC does not result from the re-expression of a fetal marker. Indeed, analysis of mouse embryos has revealed that HIP/PAP is not expressed in the liver during development [7]. HIP/PAP has also been detected in the pancreas and in a subset of cells (Paneth cells) in the intestine [8]. Moreover in rats, the HIP/PAP homologue (PAP 1/peptide 23/Reg 2), is expressed in pituitary and uterine cells under the influence of growth hormone releasing hormone and oestradiol, respectively [9,10], and by motor neurones in vivo during their regeneration and in vitro when incubated with ciliary neurotrophic factor-related cytokines [11,12].Little is known about the physiopathological significance of HIP/PAP expression. In the pancreas, there is evidence that HIP/PAP may participate in the antiapoptotic programme developed by acinar cells during acute pancreatitis [13]; indeed, HIP/PAP was reported to protect pancreatic AR4-2 J cells against apoptosis induced by oxidative stress [14]. In pituitary cells, PAP1/peptide 23 was reported to act as a growth...

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Section: Identification Of the Riia Regulatory Subunit Of Pka As A Pasupporting

confidence: 86%

HIP/PAP, a C‐type lectin overexpressed in hepatocellular carcinoma, binds the RIIα regulatory subunit of cAMP‐dependent protein kinase and alters the cAMP‐dependent protein kinase signalling

Philippe

Sar

Pileire

et al. 2004

European Journal of Biochemistry

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“…These cells retained the characteristics of duct epithelial cells, showing a cobblestone appearance under permissive culture conditions and positive for desmosomes observed by electron microscopy and E-cadherin and carbonic anhydrase II, both of which are expressed in duct epithelial cells (Blouin et al 1997;Koisumi et al 2004) observed by immunocytochemistry (Figs. 5A-D).…”

Section: Expression Of Fas In the Pancreatic Ductular Cellsmentioning

confidence: 75%

Toll-Like Receptor 3 Signaling Induces Chronic Pancreatitis through the Fas/Fas Ligand-Mediated Cytotoxicity

Soga

Komori

Miyazaki

et al. 2009

Tohoku J. Exp. Med.

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“…Light and electron microscopic immunocytochemistry experiments were carried out as described previously (Brown and Farquhar 1989; Blouin et al 1997). DLK/MUK/ZPK antiserum was used at a dilution of 1:400 for both light and electron microscopic observations.…”

Section: Methodsmentioning

confidence: 99%

Localization of the Mixed-lineage Kinase DLK/MUK/ZPK to the Golgi Apparatus in NIH 3T3 Cells

Douziech

Laberge

Grondin

et al. 1999

J Histochem Cytochem.

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SUMMARY DLK/MUK/ZPK is a serine/threonine kinase that belongs to the mixed-lineage (MLK) subfamily of protein kinases. As is the case for most members of this family, relatively little is known about the physiological role of DLK/MUK/ZPK in mammalian cells. Because analysis of subcellular distribution may provide important clues concerning the potential in vivo function of a protein, an antiserum was generated against the amino terminal region of murine DLK/MUK/ZPK and used for localization studies in wild-type NIH 3T3 cells. Light microscopic immunocytochemistry experiments performed with the antiserum revealed that DLK/MUK/ZPK was specifically localized in a juxtanuclear structure characteristic of the Golgi complex. In support of this, treatment of cells with brefeldin A, a drug known to disintegrate the Golgi apparatus, caused disruption of DLK/MUK/ZPK perinuclear staining. Ultrastructural observation of NIH 3T3 cells also confirmed this localization, showing that most of the immunoreactivity was detected on membranes of the stacked Golgi cisternae. Consistent with localization studies, biochemical analyses revealed that DLK/MUK/ZPK was predominantly associated with Golgi membranes on fractionation of cellular extracts and was entirely partitioned into the aqueous phase when membranes were subjected to Triton X-114 extraction. On the basis of these findings, we suggest that DLK/MUK/ZPK is a peripheral membrane protein tightly associated with the cytoplasmic face of the Golgi apparatus. (J Histochem Cytochem 47:1287-1296, 1999)

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Establishment and immunocharacterization of an immortalized pancreatic cell line derived from the H-2Kb-tsA58 transgenic mouse

Cited by 8 publications

References 24 publications

HIP/PAP, a C‐type lectin overexpressed in hepatocellular carcinoma, binds the RIIα regulatory subunit of cAMP‐dependent protein kinase and alters the cAMP‐dependent protein kinase signalling

HIP/PAP, a C‐type lectin overexpressed in hepatocellular carcinoma, binds the RIIα regulatory subunit of cAMP‐dependent protein kinase and alters the cAMP‐dependent protein kinase signalling

Toll-Like Receptor 3 Signaling Induces Chronic Pancreatitis through the Fas/Fas Ligand-Mediated Cytotoxicity

Localization of the Mixed-lineage Kinase DLK/MUK/ZPK to the Golgi Apparatus in NIH 3T3 Cells

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