Different response to epidermal growth factor of hepatocytes in cultures isolated from male or female rat liver

Francavilla, A.; Ove, Peter; Polimeno, Lorenzo; Sciascia, C; Coetzee, Mona L.; Pellici, Riccardo; Todo, Satoru; Kam, Igal; Starzl, Thomas E.

doi:10.1016/0016-5085(87)90924-3

Cited by 31 publications

(17 citation statements)

References 28 publications

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“…Hepatocytes were isolated by modification of the in situ two-step collagenase perfusion technique of Seglen [15], as previously described [16]. The purity of the preparation was more than 98%, as demonstrated by the staining for glucose-6-phosphat ase [16].…”

Section: Methodsmentioning

confidence: 99%

Ursodeoxycholate promotes protein phosphorylation in the cytosol of rat hepatocytes

et al. 1997

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SummaryA study is presented of the effect of the bile salt ursodeoxycholate (UDC) on protein phosphorylation by [~,-32P]ATP in the cytosol from rat hepatocytes. Gel electrophoresis and corresponding autoradiograms of cytosolic proteins show that UDC promotes phosphorylation of at least eight different protein bands. Four of them (the 36, 60, 64 and 76 kDa) are phosphorylated by Ca 2+ and phospholipid-dependent protein kinase (PKC); three (the 31, 51 and 71 kDa) are phosphorylated by cAMPdependent protein kinase (PKA) and one protein band, with molecular weight of 34 kDa, apparently contains substrates of both PKC and PKA. Data are reported indicating that UDC can directly affect the intrinsic activity of protein kinases.

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Section: Methodsmentioning

confidence: 99%

Ursodeoxycholate promotes protein phosphorylation in the cytosol of rat hepatocytes

et al. 1997

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“…The preparation of cytosolic, nuclear, and plasma membrane fractions was as described previously (10,15). The methods for the estrogen receptor assay (10) and the EGF binding assay have been described previously (15).…”

Section: Binding Studiesmentioning

confidence: 99%

“…The methods for the estrogen receptor assay (10) and the EGF binding assay have been described previously (15).…”

Section: Binding Studiesmentioning

confidence: 99%

Role of estrogens and epidermal growth factor in hepatocellular carcinoma (HCC)

et al. 1991

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Estrogen (E) and epidermal growth factors (EGF) receptors were assayed in the liver of nine patients with hepatocellular carcinoma (HCC). Total E and nuclear E receptors were decreased significantly in neoplastic tissue as compared to the levels found in surrounding nonneoplastic tissue. The EGF receptor was decreased also in neoplastic tissue. On the basis of binding data, a decrease in the number but not in affinity of both the E and EGF receptors was found. Keywordsestrogen; epidermal growth factor receptors; hepatocellular carcinoma Epidermal growth factor (EGF) is a polypeptide growth factor that was isolated originally from the submaxillary gland. It stimulates DNA synthesis in vitro in a variety of different cells (1)(2)(3). This effect is initiated by the interaction of EGF with a specific plasma membrane receptor (4). During the last five years, several lines of evidence have demonstrated that EGF receptor expression can be altered by hormones such as estrogen (E) (4), growth hormone (5), and thyroid stimulating hormone (TSH) (6) in target tissues for these hormones.The relationship between E and EGF has been studied extensively in normal uterus and in breast cancer tissue (4-7). For example, it has been shown that E stimulates EGF receptor expression in uterine tissue obtained from immature rats (4). In contrast, an inverse relationship exists between the EGF and E receptors content in breast cancer tissues (8). Recently it has been shown that the liver is also a target tissue for sex hormones (9-11). E receptors (ER) appear to play an important role in hepatic regeneration where a strong temporal relationship between increased hepatic DNA synthesis and an increased activity and nuclear distribution of ER have been found. Because both ER and EGF are involved in hepatocyte proliferation, the pattern of E and EGF receptors in tissue samples obtained from patients with hepatocellular carcinoma (HCC) was studied. MATERIALS AND METHODS Liver SpecimensSamples of HCC-containing and normal adjacent liver were collected from patients undergoing resection in Milan, Bari, and Pittsburgh. Both tumor and adjacent normal liver tissue were collected and their identities confirmed by histology. The samples were wrapped in aluminum foil, snap-frozen, and stored at −70° C until assayed for their content of E and EGF receptors. Prior studies using fresh and frozen liver have showed that both the E and EGF receptors are stable during frozen storage. MaterialsRadioactive [2,3,6, H]estradiol ([ 3 H]E 2 ), 90 Ci/mmol and [ 125 I]EGF were purchased from New England Nuclear. The purity of the radiolabeled compounds was assessed periodically by thin-layer or column chromatography (12). Sources of other materials were as described previously (12)(13)(14). Binding StudiesThe preparation of cytosolic, nuclear, and plasma membrane fractions was as described previously (10,15). The methods for the estrogen receptor assay (10) and the EGF binding assay have been described previously (15). Other MethodsProtein concentrations ...

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“…Efforts to identify and purify the active constituent in the cytosol of rat livers were hampered by the inability to settle on an appropriate assay. Fractions prepared by other investigators were reported to be mitogenic in tissue cultures of hepatocytes or hepatoma cells (12,13,15), whereas ours were inert in vitro throughout purification to the final fraction used for amino acid sequence analysis and cDNA cloning (9,10,26).…”

Section: Discussionmentioning

confidence: 68%

“…The method of purification of ALR from male Fisher weanling rats was as reported (9,10). The procedures involved the following successive steps: ethanol precipitation, ultrafiltration through an Amicon PM30 membrane, cation-exchange FPLC on a Mono Q column, and nondissociating PAGE.…”

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confidence: 99%

Cloning and sequence analysis of the rat augmenter of liver regeneration (ALR) gene: expression of biologically active recombinant ALR and demonstration of tissue distribution.

Hagiya

Francavilla

Polimeno

et al. 1994

Proc. Natl. Acad. Sci. U.S.A.

157

120

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A full-length cDNA clone encoding a purified augmenter of liver regeneration (ALR) factor prepared from the cytosol of weanling rat livers was isolated. The 1.2-kb cDNA included a 299-bp 5' untranslated region, a 375-bp coding region, and a 550-bp 3' untranslated region. It encoded a protein consisting of 125 amino acids. The molecular weight of ALR calculated from the cDNA was 15,081, which is consistent with the size estimated by SDS/PAGE under reducing conditions. The molecular weight of the purified native ALR estimated by SDS/PAGE under nonreducing conditions was -30,000; thus ALR apparently has a homodimeric structure. The recombinant ALR produced by expression of the cDNA in COS cells was tested in vivo in the canine Eck fistula model and found to have potency equivalent to the purified native ALR. The 125-aa sequence deduced from the rat ALR cDNA shows 50% homology to the amino acid sequence of the gene for oxidative phosphorylation and vegetative growth in the yeast Saccharomyces cerevisiae.

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Different response to epidermal growth factor of hepatocytes in cultures isolated from male or female rat liver

Cited by 31 publications

References 28 publications

Ursodeoxycholate promotes protein phosphorylation in the cytosol of rat hepatocytes

Ursodeoxycholate promotes protein phosphorylation in the cytosol of rat hepatocytes

Role of estrogens and epidermal growth factor in hepatocellular carcinoma (HCC)

Cloning and sequence analysis of the rat augmenter of liver regeneration (ALR) gene: expression of biologically active recombinant ALR and demonstration of tissue distribution.

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