Abstract
To construct a hepatic stellate cells (HSCs) subtracted cDNA library to find differentially expressed genes in normal mice and mice infected withSchistosoma japonicum (S. japonicum). Suppression subtractive hybridization (SSH) was used. The cDNA fragments of normal mouse were compared to those ofschistosoma-infected mice to find differentially expressed genes. Then differentially expressed cDNA fragments were directly inserted into T/A cloning vector to set up the subtractive library. Amplification of the library was carried out with transformation of DH5α. The amplified library contained more than 400 positive bacterial clones, which were then hybridized with forward and backward subtracted probes for differential screening. One hundred positive bacterial clones were randomly selected for sequencing and BLAST analysis. Finally, virtual Northern Blot confirmed such differential expression. The subtracted cDNA library of differentially expressed genes of HSCs was constructed successfully, the library is efficient and lays foundation for screening and cloning new and specific genes of schistosomiasis.
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Aikawa, R., Komuro, I., Nagai, R., Yazaki, Y., 2000. Rho plays an important role in angiotensin II-induced hypertrophic responses in cardiac myocytes.Mol Cell Biochem,212:177–182.
Cassiman, D., Denef, C., Desmet, V., Roskams, T., 2001. Human and rat hepatic stellate cells express neurotrophins and neurotrophin receptors.Hepatology,33:148–158.
Choi, S.S., Yun, J.W., Choi, E.K., Cho, Y.G., Sung, Y.C., Shin, H.S., 1995. Construction of a gene expression profile of a human fetal liver by single-pass cDNA sequencing.Mamm-Genome,6:653–657.
Cintin, C., Johansen, J.S., Skov, F., Price, P.A., Nielsen, H.J., 2001. Accumulation of the neutrophil-derived protein YKL-40 during storage of various blood components.Inflammation Research,50:107–111.
Diatchenko, L., Lau, Y.C., Campbell, A.P., Chenchik, A., Moqadam, F., Huang, B., Lukyanov, S., Lukyanov, K., Gurskaya, N., Sverdlov, E.D., Siebert, P.D., 1996. Suppression subtractive hybridization: A method for generating differentially regulated or tissue-specific cDNA probes and libraries.Proc Natl Acid Sci USA,93:6025–6030.
Diaz, A., Willis, A.C., Sim, R.B., 2000. Expression of the proteinase specialized in bone resorption, cathepsin K, in granulomatous inflammation.Mol Med,6(8):648–659.
Di Sario, A., Bendia, E., Svegliati-Baroni, G., Marzioni, M., Ridolfi, F., Trozzi, L., Ugili, L., Saccomanno, S., Jezequel, A.M., Benedetti, A., 2002. Rearrangement of the cytoskeletal network induced by platelet-derived growth factor in rat hepatic stellate cells: role of different intracellular signalling pathways.J Hepatol,36:179–190.
Dodds, R.A., James, I.E., Rieman, D., Ahern, R., Hwang, S.M., Connor, J.R., Thompson, S.D., Veber, D.F., Drake, F.H., Holmes, S., Lark, M.W., Gowen, M., 2001. Human osteoclast cathepsin K is processed intracellularly prior to attachment and bone resorption.J Bone Miner Res,16:478–486.
Drew, A.C., Brindley, P.J., 1995. Female-specific sequences isolated from Schistosoma mansoni by representational difference analysis.Mol Biochem Parasitol,71:173–181.
Franco, G.R., Rabelo, E.M., Azevedo, V., 1997. Evaluation of cDNA libraries from different developmental stages of Schistosoma mansoni for production of expressed sequence tags (ESTs).DNA-Res,4:231–240.
Friedman, S.L., 1993. Seminars in medicine of the Beth Israel Hospital, Boston. The cellular basis of hepatic fibrosis. Mechanisms and treatment strategies.N Engl J Med,328:1828–1835.
Hakala, B.E., White, C., Recklies, A.D., 1993. Human cartilage gp39, a major secretory product of articular chondrocytes and synovial cells, is a mammalian member of a chitinase protein family.J Biol Chem,268:25803–25810.
Johansen, J.S., Williamson, M.K., Rice, J.S., Price, P.A., 1992. Identification of proteins secreted by human osteoblastic cells in culture.J Bone Miner Res,7:501–512.
Johansen, J.S., Christoffersen, P., Moller, S., Price, P.A., Henriksen, J.H., Garbarsch, C., Bendtsen, F., 2000. Serum YKL-40 is increased in patients with hepatic fibrosis.J Hepatol,32(6):911–920.
Kowalczyk, P., Woszczynski, M., Ostrowski, J., 2002. Increased expression of ribosomal protein S2 in liver tumors, posthepactomized livers, and proliferating hepatocytes in vitro.Acta Biochim Pol,49:615–624.
Kuang, W.W., Thompson, D.A., Hoch, R.V., Weigel, R.J., 1998. Differential screening and suppression subtractive hybridization identified genes differentially expressed in an estrogen receptor-positive breast carcinoma cell line.Nucleic Acids Res,26:1116–1123.
Kwon, K.B., Park, E.K., Ryu, D.G., Park, B.H., 2002. D4-GDI is cleaved by caspase-3 during daunorubicin-induced apoptosis in HL-60 cells.Exp Mol Med,34:32–37.
Lee, J.H., Miele, M.E., Hicks, D.J., Phillips, K.K., Trent, J.M., Weissman, B.E., Welch, D.R., 1996. KISS-I, a novel human malignant melanoma metastasis-suppressor gene.J Natl Cancer Inst,88:1731–1737.
Levy, M.T., McCaughan, G.W., Abbott, C.A., Park, J.E., Cunningham, A.M., Muller, E., Rettig, W.J., Gorrell, M.D., 1999. Fibroblast activation protein: a cell surface dipeptidyl peptidase and gelatinase expressed by stellate cells at the tissue remodeling interface in human cirrhosis.Hepatology,29:1768–1778.
Niki, T., Pekny, M., Hellemans, K., Bleser, P.D., Berg, K.V., Vaeyens, F., Quartier, E., Schuit, F., Geerts, A., 1999. Class IV intermediate filament protein nestin is induced during activation of rat hepatic stellate cells.Hepatology,29:520–527.
Nojgaard, C., Johansen, J.S., Christensen, E., Skovgaard, L.T., Price, P.A., Becker, U., 2003a. Serum levels of YKL-40 and PIIINP as prognostic markers in patients with alcoholic liver disease.J Hepatol,39:179–186.
Nojgaard, C., Johansen, J.S., Krarup, H.B., Holten-Andersen, M., Moller, A., Bendtsen, F., 2003b. Effect of antiviral therapy on markers of fibrogenesis in patients with chronic hepatitis C.Scand J Gastroenterol,38:659–665.
Nyirkos, P., Golds, E.E., 1990. Human synovial cells secrete a 39 kDa protein similar to a bovine mammary protein expressed during the non-lactating period.Biochem J,269:265–268.
Pettersson, M., Dannaeus, K., Nilsson, K., Jonsson, J.I., 2000. Isolation of MYADM, a novel hematopoietic-associated marker gene expressed in multipotent progenitor cells and up-regulated during myeloid differentiation.J Leukoc Biol,67:423–431.
Pinzani, M., 1995. Novel insights into the biology and physiology of the Ito cell.Pharmacol Ther,66:387–412.
Pinzani, M., 2002. PDGF and signal transduction in hepatic stellate cells.Front Biosci,7:d1720-d1726.
Pitzer, C., Stassar, M., Zoller, M., 1999. Identification of renal-cell-carcinoma-related cDNA clones by suppression subtractive hybridization.J Cancer Res Clin Oncol,125:487–492.
Recklies, A.D., White, C., Ling, H., 2002. The chitinase 3-like protein human cartilage glycoprotein 39 (HC-gp39) stimulates proliferation of human connective-tissue cells and activates both extracellular signal-regulated kinase-and protein kinase B-mediated signalling pathways.Biochem J,365:119–126.
Reis, M.G., Kuhns, J., Blanton, R., Davis, A.H., 1989. Localization and pattern of expression of a female specific mRNA in Schistosoma mansoni.Mol Biochem Parasitol,32:113–119.
Renkema, G.H., Boot, R.G., Au, F.L., Donker-Koopman, W.E., Strijland, A., Muijsers, A.O., Hrebicek, M., Aerts, J.M., 1998. Chitotriosidase, a chitinase, and the 39-kDa human cartilage glycoprotein, a chitin-binding lectin, are homologues of family 18 glycosyl hydrolases secreted by human macrophages.Eur J Biochem,251:504–509.
Shackel, N.A., McGuinness, P.H., Abbott, C.A., Gorrell, M.D., McCaughan, G.W., 2003. Novel differential gene expression in human cirrhosis detected by suppression subtractive hybridization.Hepatology,38:577–588.
Sorensen, E., Bogh, H.O., Johansen, M.V., McManus, D.P., 1999. PCR-based identification of individuals of Schistosoma japonicum representing different subpopulations using a genetic marker in mitochondrial DNA.Int J Parasitol,29:1121–1128.
Tada, S., Iwamoto, H., Nakamuta, M., Sugimoto, R., Enjoji, M., Nakashima, Y., Nawata, H., 2001. A selective ROCK inhibitor, Y27632, prevents dimethylnitrosamine-induced hepatic fibrosis in rats.J Hepatol,34:529–536.
Volck, B., Price, P.A., Johansen, J.S., Sorensen, O., Benfield, T.L., Nielsen, H.J., Calafat, J., Borregaard, N., 1998. YKL-40, a mammalian member of the chitinase family, is a matrix protein of specific granules in human neutrophils.Proc Assoc Am Physicians,110:351–360.
Von Stein, O.D., Thies, W.G., Hofmann, M., 1997. A high throughout screening for rarely transcribed differentially expression genes.Nucleic Acids Res,25:2598–2602.
Zhang, L., Cilley, R.E., Chinoy, M.R., 2000. Suppression subtractive hybridization to identify gene expressions in variant and classic small cell lung cancer cell lines.J Surg Res,93:108–119.
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Project (No. 2001BA705B08) supported by the National Ten-year Key Technologies R&D Program, China
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Min, Z., Yi-jun, W., Wei-min, C. et al. Construction of a hepatic stellate cells subtracted cDNA library of differentially expressed genes in normal mice and mice with Schistosomiasis japonica. J Zheijang Univ Sci B 6, 280–287 (2005). https://doi.org/10.1007/BF02842465
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DOI: https://doi.org/10.1007/BF02842465