Abstract
Culture of Hep G2 cells in medium containing 2% (v/v) dimethyl sulphoxide (DMSO) resulted in a slowing of growth and a marked change in morphological appearance. By day 6, cultures containing DMSO had one-third the number of cells compared with parallel control cultures. Measurement of 125I-epidermal-growth-factor (EGF) binding to DMSO-treated cells revealed a striking time-dependent elevation in specific EGF binding to their cell surface. Increased binding was detectable within 24 h of the start of DMSO treatment, reaching, by 6 days, levels almost 25 times greater than those for control cells. Addition of EGF to DMSO-treated cells caused a rapid down-regulation of the EGF receptor, but did not alter their proliferation rate. Slowing of growth by other means, such as serum starvation, growth to confluence or culture in the presence of sodium butyrate, did not affect 125I-EGF binding, indicating a specific effect of DMSO on these cells.
Full text
PDF




Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Ben-Ze'ev A., Robinson G. S., Bucher N. L., Farmer S. R. Cell-cell and cell-matrix interactions differentially regulate the expression of hepatic and cytoskeletal genes in primary cultures of rat hepatocytes. Proc Natl Acad Sci U S A. 1988 Apr;85(7):2161–2165. doi: 10.1073/pnas.85.7.2161. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Berry M. N., Friend D. S. High-yield preparation of isolated rat liver parenchymal cells: a biochemical and fine structural study. J Cell Biol. 1969 Dec;43(3):506–520. doi: 10.1083/jcb.43.3.506. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Carlin C. R., Simon D., Mattison J., Knowles B. B. Expression and biosynthetic variation of the epidermal growth factor receptor in human hepatocellular carcinoma-derived cell lines. Mol Cell Biol. 1988 Jan;8(1):25–34. doi: 10.1128/mcb.8.1.25. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chan K., Kost D. P., Michalopoulos G. Multiple sequential periods of DNA synthesis and quiescence in primary hepatocyte cultures maintained on the DMSO-EGF on/off protocol. J Cell Physiol. 1989 Dec;141(3):584–590. doi: 10.1002/jcp.1041410317. [DOI] [PubMed] [Google Scholar]
- Clementi M., Festa A., Testa I., Bagnarelli P., Devescovi G., Carloni G. Expression of high- and low-affinity epidermal growth factor receptors in human hepatoma cell lines. FEBS Lett. 1989 Jun 5;249(2):297–301. doi: 10.1016/0014-5793(89)80645-3. [DOI] [PubMed] [Google Scholar]
- Earp H. S., Lin Q., Blaisdell J. Dimethyl sulfoxide decreases specific EGF binding. J Cell Biochem. 1984;26(4):221–230. doi: 10.1002/jcb.240260403. [DOI] [PubMed] [Google Scholar]
- Earp H. S., O'Keefe E. J. Epidermal growth factor receptor number decreases during rat liver regeneration. J Clin Invest. 1981 May;67(5):1580–1583. doi: 10.1172/JCI110190. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Francavilla A., Ove P., Polimeno L., Sciascia C., Coetzee M. L., Starzl T. E. Epidermal growth factor and proliferation in rat hepatocytes in primary culture isolated at different times after partial hepatectomy. Cancer Res. 1986 Mar;46(3):1318–1323. [PMC free article] [PubMed] [Google Scholar]
- Galbraith R. A., Sassa S., Kappas A. Induction of haem synthesis in Hep G2 human hepatoma cells by dimethyl sulphoxide. A transcriptionally activated event. Biochem J. 1986 Jul 15;237(2):597–600. doi: 10.1042/bj2370597. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gladhaug I. P., Refsnes M., Christoffersen T. Regulation of hepatocyte epidermal growth factor receptors by n-butyrate and dimethyl sulfoxide: sensitivity to modulation by the tumor promoter TPA. Anticancer Res. 1989 Nov-Dec;9(6):1587–1592. [PubMed] [Google Scholar]
- Gladhaug I. P., Refsnes M., Sand T. E., Christoffersen T. Effects of butyrate on epidermal growth factor receptor binding, morphology, and DNA synthesis in cultured rat hepatocytes. Cancer Res. 1988 Nov 15;48(22):6560–6564. [PubMed] [Google Scholar]
- HUNTER W. M., GREENWOOD F. C. Preparation of iodine-131 labelled human growth hormone of high specific activity. Nature. 1962 May 5;194:495–496. doi: 10.1038/194495a0. [DOI] [PubMed] [Google Scholar]
- Hatada E. N., McClain D. A., Potter E., Ullrich A., Olefsky J. M. Effects of growth and insulin treatment on the levels of insulin receptors and their mRNA in Hep G2 cells. J Biol Chem. 1989 Apr 25;264(12):6741–6747. [PubMed] [Google Scholar]
- Isom H. C., Secott T., Georgoff I., Woodworth C., Mummaw J. Maintenance of differentiated rat hepatocytes in primary culture. Proc Natl Acad Sci U S A. 1985 May;82(10):3252–3256. doi: 10.1073/pnas.82.10.3252. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kan M., Huang J. S., Mansson P. E., Yasumitsu H., Carr B., McKeehan W. L. Heparin-binding growth factor type 1 (acidic fibroblast growth factor): a potential biphasic autocrine and paracrine regulator of hepatocyte regeneration. Proc Natl Acad Sci U S A. 1989 Oct;86(19):7432–7436. doi: 10.1073/pnas.86.19.7432. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kapuściński J., Skoczylas B. Simple and rapid fluorimetric method for DNA microassay. Anal Biochem. 1977 Nov;83(1):252–257. doi: 10.1016/0003-2697(77)90533-4. [DOI] [PubMed] [Google Scholar]
- Kelly J. H., Darlington G. J. Modulation of the liver specific phenotype in the human hepatoblastoma line Hep G2. In Vitro Cell Dev Biol. 1989 Feb;25(2):217–222. doi: 10.1007/BF02626182. [DOI] [PubMed] [Google Scholar]
- Knowles B. B., Howe C. C., Aden D. P. Human hepatocellular carcinoma cell lines secrete the major plasma proteins and hepatitis B surface antigen. Science. 1980 Jul 25;209(4455):497–499. doi: 10.1126/science.6248960. [DOI] [PubMed] [Google Scholar]
- Kuppuswamy D., Pike L. J. Ligand-induced desensitization of 125I-epidermal growth factor internalization. J Biol Chem. 1989 Feb 25;264(6):3357–3363. [PubMed] [Google Scholar]
- McGowan J. A. Reciprocal regulation of adult rat hepatocyte growth and functional activities in culture by dimethyl sulfoxide. J Cell Physiol. 1988 Dec;137(3):497–504. doi: 10.1002/jcp.1041370315. [DOI] [PubMed] [Google Scholar]
- Michalopoulos G., Cianciulli H. D., Novotny A. R., Kligerman A. D., Strom S. C., Jirtle R. L. Liver regeneration studies with rat hepatocytes in primary culture. Cancer Res. 1982 Nov;42(11):4673–4682. [PubMed] [Google Scholar]
- Miyazaki M., Handa Y., Oda M., Yabe T., Miyano K., Sato J. Long-term survival of functional hepatocytes from adult rat in the presence of phenobarbital in primary culture. Exp Cell Res. 1985 Jul;159(1):176–190. doi: 10.1016/s0014-4827(85)80047-1. [DOI] [PubMed] [Google Scholar]
- Nakamura T., Tomita Y., Ichihara A. Density-dependent growth control of adult rat hepatocytes in primary culture. J Biochem. 1983 Oct;94(4):1029–1035. doi: 10.1093/oxfordjournals.jbchem.a134444. [DOI] [PubMed] [Google Scholar]
- Nakamura T., Yoshimoto K., Nakayama Y., Tomita Y., Ichihara A. Reciprocal modulation of growth and differentiated functions of mature rat hepatocytes in primary culture by cell--cell contact and cell membranes. Proc Natl Acad Sci U S A. 1983 Dec;80(23):7229–7233. doi: 10.1073/pnas.80.23.7229. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Savage C. R., Jr, Cohen S. Epidermal growth factor and a new derivative. Rapid isolation procedures and biological and chemical characterization. J Biol Chem. 1972 Dec 10;247(23):7609–7611. [PubMed] [Google Scholar]
- Schacterle G. R., Pollack R. L. A simplified method for the quantitative assay of small amounts of protein in biologic material. Anal Biochem. 1973 Feb;51(2):654–655. doi: 10.1016/0003-2697(73)90523-x. [DOI] [PubMed] [Google Scholar]
- Selden C., Hodgson H. J. Further characterisation of 'hepatotropin', a high molecular weight hepatotrophic factor in rat serum. J Hepatol. 1989 Sep;9(2):167–176. doi: 10.1016/0168-8278(89)90047-0. [DOI] [PubMed] [Google Scholar]
- Skov Olsen P., Boesby S., Kirkegaard P., Therkelsen K., Almdal T., Poulsen S. S., Nexø E. Influence of epidermal growth factor on liver regeneration after partial hepatectomy in rats. Hepatology. 1988 Sep-Oct;8(5):992–996. doi: 10.1002/hep.1840080503. [DOI] [PubMed] [Google Scholar]

