Abstract
Aim:
Overexpression of midkine (MK) has been observed in many malignancies. This aim of this study is to screen for suitable antisense oligonucleotides (ASODN) targeting MK in hepatocellular carcinoma (HCC) cells and evaluate its antitumor activity.
Methods:
Ten ASODN targeting MK were designed and synthesized. After transfection with ASODN, cell proliferation was analyzed with MTS [3 -(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt] assay. In addition, MK mRNA, protein levels, as well as apoptosis and caspase-3 activity were also examined in HepG2 cells. Cell proliferation was then analyzed after treatment with both ASODN and chemotherapeutic drugs.
Results:
In this experiment, the ASODN5 among the 10 ASODN showed higher inhibitory activity against proliferation of hepatocellular carcinoma cells in a dose-dependent manner. In HepG2 cells, ASODN5 could significantly reduce the MK mRNA level and protein content. After transfection with ASODN5 for 48 h, accompanied with a decline of survivin and Bcl-2 protein content, a remarkable increase of apoptosis and caspase-3 activity was observed in HepG2 cells. Furthermore, ASODN5 transfer can significantly increase chemosensitivity in HepG2 cells.
Conclusion:
Antisense oligonucleotides targeting MK shows therapeutic effects on HCC; ASODN5 has the possibility to be developed as an effective antitumor agent.
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Kadomatsu K, Tomomura M, Muramatsu T . cDNA cloning and sequencing of a new gene intensely expressed in early differentiation stages of embryonal carcinoma cells and in mid-gestation period of mouse embryogenesis. Biochem Biophys Res Commun 1988; 151: 1312–8.
Li YS, Milner PG, Chauhan AK, Watson MA, Hoffman RM, Kodner CM, et al. Cloning and expression of a developmentally regulated protein that induces mitogenic and neurite outgrowth activity. Science 1990; 250: 1690–4.
Merenmies J, Rauvala H . Molecular cloning of the 18-kDa growth-associated protein of developing brain. J Biol Chem 1990; 265: 16721–4.
Muramatsu T . Midkine (MK), the product of a retinoic acid responsive gene, and pleiotrophin constitute a new protein family regulating growth and differentiation. Int J Dev Biol 1993; 37: 183–8.
Garver RI, Chan CS, Milner PG . Reciprocal expression of pleiotrophin and midkine in normal versus malignant lung tissues. Am J Respir Cell Mol Biol 1993; 9: 463–6.
Garver RI, Radford DM, Donis-Keller H, Wick MR, Milner PG . Midkine and pleiotrophin expression in normal and malignant breast tissue. Cancer 1994; 74: 1584–90.
Aridome K, Tsutsui J, Takao S, Kadomatsu K, Ozawa M, Aikou T . Increased midkine gene expression in human gastrointestinal cancers. Jpn J Cancer Res 1995; 86: 655–61.
Nakanishi T, Kadomatsu K, Okamoto T, Tomoda Y, Muramatsu T . Expression of midkine and pleiotropin in ovarian tumors. Obstet Gynecol 1997; 90: 285–90.
O'Brien T, Cranston D, Fuggle S, Bicknell R, Harris AL . The angiogenic factor midkine is expressed in bladder cancer, and overexpression correlates with a poor outcome in patients with invasive cancers. Cancer Res 1996; 56: 2515–8.
Konishi N, Nakamura M, Nakaoka S, Hiasa Y, Cho M, Uemura H, et al. Immuno-histochemical analysis of midkine expression in human prostate carcinoma. Oncology 1999; 57: 253–7.
Mishima K, Asai A, Kadomatsu K, Ino Y, Nomura K, Narita Y, et al. Increased expression of midkine during the progression of human astrocytomas. Neurosci Lett 1997; 233: 29–32.
Nakagawara A, Milbrandt J, Muramatsu T, Deuel TF, Zhao H, Cnaan A, et al. Differential expression of pleiotrophin and midkine in advanced neuroblastomas. Cancer Res 1995; 55: 1792–7.
Tsutsui J, Kadomatsu K, Matsubara S, Nakagawara A, Hamanoue M, Takao S, et al. A new family of heparin-binding growth/differentiation factors: increased midkine expression in Wilms' tumor and other human carcinomas. Cancer Res 1993; 53: 1281–5.
Li YS, Milner PG, Chauhan AK, Watson MA, Hoffman RM, Kodner CM, et al. Cloning and expression of a developmentally regulated protein that induces mitogenic and neurite outgrowth activity. Science 1990; 250: 1690–4.
Shimada H, Nabeya Y, Tagawa M, Okazumi S, Matsubara H, Okazumi S, et al. Preoperative serum midkine concentration is a prognostic marker for esophageal squamous cell carcinoma. Cancer Sci 2003; 94: 628–32.
Ikematsu S, Nakagawara A, Nakamura Y, Sakuma S, Wakai K, Muramatsu T, et al. Correlation of elevated level of blood midkine with poor prognostic factors of human neuroblastomas. Br J Cancer 2003; 88: 1522–6.
Obata Y, Kikuchi S, Lin Y, Yagyu K, Muramatsu T, Kumai H, et al. Serum midkine concentrations and gastric cancer. Cancer Sci 2005; 96: 54–6.
Owada K, Sanjo N, Kobayashi T, Mizusawa H, Muramatsu H, Muramatsu T, et al. Midkine inhibits caspase-dependent apoptosis via the activation of mitogen-activated protein kinase and phosphatidylinositol 3-kinase in cultured neurons. J Neurochem 1999; 73: 2084–92.
Qi M, Ikematsu S, Ichihara-Tanaka K, Sakuma S, Muramatsu T, Kadomatsu K . Midkine rescues Wilms' tumor cells from cisplatin-induced apoptosis: regulation of Bcl-2 expression by midkine. J Biochem 2000; 127: 269–77.
Muramatsu H, Muramatsu T . Purification of recombinant midkine and examination of its biological activities: functional comparison of new heparin binding factors. Biochem Biophys Res Commun 1991; 177: 652–8.
Muramatsu H, Shirahama H, Yonezawa S, Maruta H, Muramatsu T . Midkine (MK), a retinoic acid-inducible growth/differentiation factor: immunochemical evidence for the function and distribution. Dev Biol 1993; 159: 392–402.
Takada T, Toriyama K, Muramatsu H, Song XJ, Torii S, Muramatsu T . Midkine, a retinoic acid-inducible heparin-binding cytokine in inflammatory responses: chemotactic activity to neutrophils and association with inflammatory synovitis. J Biochem 1997; 122: 453–8.
Maeda N, Ichihara-Tanaka K, Kimura T, Kadomatsu K, Muramatsu T, Noda M . A receptor-like protein-tyrosine phosphatase PTPzeta/RPTPbeta binds a heparin-binding growth factor midkine. Involvement of arginine 78 of midkine in the high affinity binding to PTPzeta. J Biol Chem 1999; 274: 12474–9.
Horiba M, Kadomatsu K, Nakamura E, Muramatsu H, Ikematsu S, Sakuma S, et al. Neointima formation in a restenosis model is suppressed in midkine-deficient mice. J Clin Invest 2000; 105: 489–95.
Sandra F, Harada H, Nakamura N, Ohishi M . Midkine induced growth of ameloblastoma through MAPK and Akt pathways. Oral Oncol 2004; 40: 274–80.
Hsu IC, Metcalf RA, Sun T, Welsh JA, Wang NJ, Harris CC . Mutational hotspot in the p53 gene in human hepatocellular carcinomas. Nature 1991; 350: 427–8.
Moriya K, Fujie H, Shintani Y, Yotsuyanagi H, Tsutsumi T, Ishibashi K, et al. The core protein of hepatitis C virus induces hepatocellular carcinoma in transgenic mice. Nat Med 1998; 4: 1065–7.
Thorgeirsson SS, Teramoto T, Factor VM . Dysregulation of apoptosis in hepatocellular carcinoma. Semin Liver Dis 1998; 18: 115–22.
Lau SH, Guan XY . Cytogenetic and molecular genetic alterations in hepatocellular carcinoma. Acta Pharmacol Sin 2005; 26: 659–65.
Park YN, Chae KJ, Kim YB, Park C, Theise N . Apoptosis and proliferation in hepatocarcinogenesis related to cirrhosis. Cancer 2001; 92: 2733–8.
Kato M, Shinozawa T, Kato S, Awaya A, Terada T . Increased midkine expression in hepatocellular carcinoma. Arch Pathol Lab Med 2000; 124: 848–52.
Dai JM, Zhang SQ, Zhang W, Lin RX, Ji ZZ, Wang SQ . Antisense oligodeoxynucleotides targeting the serine/threonine kinase Pim-2 inhibited proliferation of DU-145 cells. Acta Pharmacol Sin 2005; 26: 364–8.
Chalk AM, Sonnhammer EL . Computational antisense oligo prediction with a neural network model. Bioinformatics 2002; 18: 1567–75.
Ohuchida T, Okamoto K, Akahane K, Higure A, Todoroki H, Abe Y, et al. Midkine protects hepatocellular carcinoma cells against TRAIL-mediated apoptosis through down-regulation of caspase-3 activity. Cancer 2004; 100: 2430–6.
Jiang QS, Wang SQ . Design and screening of antisense oligodeoxynucleotides against PAI-1 mRNA in endothelial cells in vitro. Acta Pharmacol Sin 2006; 27: 1018–23.
Zuker M . Mfold web server for nucleic acid folding and hybridization prediction. Nucleic Acids Res 2003; 31: 3406–15.
Shin S, Sung BJ, Cho YS, Kim HJ, Ha NC, Hwang JI, et al. An anti-apoptotic protein human survivin is a direct inhibitor of caspase-3 and -7. Biochemistry 2001; 40: 1117–23.
Yin C, Knudson CM, Korsmeyer SJ, Van Dyke T . Bax suppresses tumorigenesis and stimulates apoptosis in vivo. Nature 1997; 385: 637–40.
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Project supported by the Zhejiang Province Medicine and Sanitation Research Foundation (2003A077).
The Editor has retracted this article because some of the figures have errors which were introduced during the process of integrating and editing the original images (Figures 3, 4, and 5). Due to the fact that the research was conducted over ten years ago, the original data can no longer be found, and the article cannot be corrected. All authors agree to this retraction.
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Dai, Lc., Wang, X., Yao, X. et al. RETRACTED ARTICLE: Antisense oligonucleotides targeting midkine induced apoptosis and increased chemosensitivity in hepatocellular carcinoma cells. Acta Pharmacol Sin 27, 1630–1636 (2006). https://doi.org/10.1111/j.1745-7254.2006.00459.x
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DOI: https://doi.org/10.1111/j.1745-7254.2006.00459.x
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