IGF expression in HPV-related and HPV-unrelated human cancer cells

Durzyńska,Julia; Barton,Elisabeth

doi:10.3892/or.2014.3329

IGF expression in HPV-related and HPV-unrelated human cancer cells

Authors:
- Julia Durzyńska
- Elisabeth Barton
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Affiliations: Department of Molecular Virology, Faculty of Biology, Institute of Experimental Biology, Adam Mickiewicz University, 61‑614 Poznań, Poland, Department of Anatomy and Cell Biology, School of Dental Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA
Published online on: July 11, 2014 https://doi.org/10.3892/or.2014.3329
Pages: 893-900
Copyright: © Durzyńska et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY_NC 3.0].

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Abstract

The human Igf-1 gene not only produces insulin‑like growth factor-I (IGF-I), but also different carboxy‑terminal extensions, known as E peptides, through alternative splicing. We and others have shown that human Eb peptide (hEb) derived from Igf-1 has intrinsic biological activity and is localized to nuclei of transfected cells. Since hEb actions can complement the activity of IGF-I itself, the aim of the present study was to compare IGF-I isoforms at the endogenous protein and transcript level in cancer cell lines, including HeLa, U2OS, HepG2 and K562 cells. Quantitative real-time PCR (qRT‑PCR) using Igf-1 isoform specific primers was performed to determine expression patterns, using β-actin as a reference gene. The overall relative Igf-1 transcript level was different across the cell lines, with ~80-fold higher expression in K562 (130.2±31.2) than in U2OS cells (1.7±1.1). The relative copy number of Igf-1b was the highest in HepG2 (69.9±28.6) and K562 cells (28.3±6.7), whereas the relative copy numbers of Igf-1a and Igf-1c were significantly higher in K562 cells compared to all other cell lines. Immunoblotting using total cell lysates, cytoplasmic and nuclear fractions were carried out to determine the level and distribution of IGF-I proteins. K562 cells exhibited the highest level of hEb in total cell lysates and nuclear fractions and no cell lines displayed hEb in the cytoplasmic fractions. In contrast, IGF-IA was the highest in HeLa cells and was enriched only in the cytoplasmic fraction. Since relatively low IGF-1A transcript level but relatively high pro‑IGF-1A protein level is plausible, we hypothesized that these transcripts could be processed with higher efficiency and/or the protein product may be stabilized by viral HPV oncogenes in HeLa cells. We assert that while it is important to analyze Igf-1 transcript level, it may be more relevant to determine the IGF isoforms at the protein level.

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1	Brett D, Pospisil H, Valcárcel J, Reich J and Bork P: Alternative splicing and genome complexity. Nat Genet. 30:29–30. 2002. View Article : Google Scholar : PubMed/NCBI
2	Roy B, Haupt LM and Griffiths LR: Review: alternative splicing (AS) of genes as an approach for generating protein complexity. Curr Genomics. 14:182–194. 2013. View Article : Google Scholar : PubMed/NCBI
3	Renaville R, Hammadi M and Portetelle D: Role of the somatotropic axis in the mammalian metabolism. Domest Anim Endocrinol. 23:351–360. 2002. View Article : Google Scholar : PubMed/NCBI
4	Langdahl BL, Kassem M, Møller MK and Eriksen EF: The effects of IGF-I and IGF-II on proliferation and differentiation of human osteoblasts and interactions with growth hormone. Eur J Clin Invest. 28:176–183. 1998. View Article : Google Scholar : PubMed/NCBI
5	Siegfried JM, Kasprzyk PG, Treston AM, Mulshine JL, Quinn KA and Cuttitta F: A mitogenic peptide amide encoded within the E peptide domain of the insulin-like growth factor IB prohormone. Proc Natl Acad Sci USA. 89:8107–8111. 1992. View Article : Google Scholar : PubMed/NCBI
6	Kuo YH and Chen TT: Novel activities of pro-IGF-I E peptides: regulation of morphological differentiation and anchorage-independent growth in human neuroblastoma cells. Exp Cell Res. 280:75–89. 2002. View Article : Google Scholar
7	Matheny RW Jr and Nindl BC: Loss of IGF-IEa or IGF-IEb impairs myogenic differentiation. Endocrinology. 152:1923–1934. 2011. View Article : Google Scholar : PubMed/NCBI
8	Hede MS, Salimova E, Piszczek A, et al: E-peptides control bioavailability of IGF-1. PLoS One. 7:e511522012. View Article : Google Scholar : PubMed/NCBI
9	Barton ER: The ABCs of IGF-I isoforms: impact on muscle hypertrophy and implications for repair. Appl Physiol Nutr Metab. 31:791–797. 2006. View Article : Google Scholar : PubMed/NCBI
10	Tian XC, Chen MJ, Pantschenko AG, Yang TJ and Chen TT: Recombinant E-peptides of pro-IGF-I have mitogenic activity. Endocrinology. 140:3387–3390. 1999. View Article : Google Scholar : PubMed/NCBI
11	Brisson BK and Barton ER: New modulators for IGF-I activity within IGF-I processing products. Front Endocrinol (Lausanne). 4:422013.PubMed/NCBI
12	Durzyńska J, Philippou A, Brisson BK, Nguyen-McCarty M and Barton ER: The pro-forms of insulin-like growth factor I (IGF-I) are predominant in skeletal muscle and alter IGF-I receptor activation. Endocrinology. 154:1215–1224. 2013.PubMed/NCBI
13	Tan DS, Cook A and Chew SL: Nucleolar localization of an isoform of the IGF-I precursor. BMC Cell Biol. 3:172002. View Article : Google Scholar : PubMed/NCBI
14	Durzyńska J, Wardziński A, Koczorowska M, Goździcka-Józefiak A and Barton ER: Human Eb peptide: not just a by-product of pre-pro-IGF1b processing? Horm Metab Res. 45:415–422. 2013.PubMed/NCBI
15	Brisson BK and Barton ER: Insulin-like growth factor-I E-peptide activity is dependent on the IGF-I receptor. PLoS One. 7:e455882012. View Article : Google Scholar : PubMed/NCBI
16	Aro AL, Savikko J, Pulkkinen V and von Willebrand E: Expression of insulin-like growth factors IGF-I and IGF-II, and their receptors during the growth and megakaryocytic differentiation of K562 cells. Leuk Res. 26:831–837. 2002. View Article : Google Scholar : PubMed/NCBI
17	Wahner Hendrickson AE, Haluska P, Schneider PA, et al: Expression of insulin receptor isoform A and insulin-like growth factor-1 receptor in human acute myelogenous leukemia: effect of the dual-receptor inhibitor BMS-536924 in vitro. Cancer Res. 69:7635–7643. 2009.PubMed/NCBI
18	Philippou A, Stavropoulou A, Sourla A, et al: Characterization of a rabbit antihuman mechano growth factor (MGF) polyclonal antibody against the last 24 amino acids of the E domain. In Vivo; 22:27–35. 2008.PubMed/NCBI
19	Harris JR and Markl J: Keyhole limpet hemocyanin (KLH): a biomedical review. Micron. 30:597–623. 1999. View Article : Google Scholar : PubMed/NCBI
20	Lowe WL Jr, Lasky SR, LeRoith D and Roberts CT Jr: Distribution and regulation of rat insulin-like growth factor I messenger ribonucleic acids encoding alternative carboxyterminal E-peptides: evidence for differential processing and regulation in liver. Mol Endocrinol. 2:528–535. 1988. View Article : Google Scholar
21	Wallis M: New insulin-like growth factor (IGF)-precursor sequences from mammalian genomes: the molecular evolution of IGFs and associated peptides in primates. Growth Horm IGF Res. 19:12–23. 2009. View Article : Google Scholar : PubMed/NCBI
22	Pfeffer LA, Brisson BK, Lei H and Barton ER: The insulin-like growth factor (IGF)-I E-peptides modulate cell entry of the mature IGF-I protein. Mol Biol Cell. 20:3810–3817. 2009. View Article : Google Scholar : PubMed/NCBI
23	Adamo ML, Neuenschwander S, LeRoith D and Roberts CT Jr: Structure, expression, and regulation of the IGF-I gene. Adv Exp Med Biol. 343:1–11. 1993. View Article : Google Scholar : PubMed/NCBI
24	Kasprzak A, Szaflarski W, Szmeja J, et al: Differential expression of IGF-1 mRNA isoforms in colorectal carcinoma and normal colon tissue. Int J Oncol. 42:305–316. 2013.PubMed/NCBI
25	Kasprzak A, Szaflarski W, Szmeja J, et al: Expression of various insulin-like growth factor-1 mRNA isoforms in colorectal cancer. Contemp Oncol (Pozn). 16:147–153. 2012.PubMed/NCBI
26	Koczorowska MM, Kwasniewska A and Gozdzicka-Jozefiak A: IGF1 mRNA isoform expression in the cervix of HPV-positive women with pre-cancerous and cancer lesions. Exp Ther Med. 2:149–156. 2011.PubMed/NCBI
27	Cui H, Yi Q, Feng J, Yang L and Tang L: Mechano growth factor E peptide regulates migration and differentiation of bone marrow mesenchymal stem cells. J Mol Endocrinol. 52:111–120. 2014. View Article : Google Scholar : PubMed/NCBI
28	Wu J, Wu K, Lin F, et al: Mechano-growth factor induces migration of rat mesenchymal stem cells by altering its mechanical properties and activating ERK pathway. Biochem Biophys Res Commun. 441:202–207. 2013. View Article : Google Scholar : PubMed/NCBI
29	Fornaro M, Hinken AC, Needle S, et al: Mechano-growth factor peptide, the COOH terminus of unprocessed insulin-like growth factor 1, has no apparent effect on myoblasts or primary muscle stem cells. Am J Physiol Endocrinol Metab. 306:E150–E156. 2014. View Article : Google Scholar : PubMed/NCBI
30	Armakolas A, Philippou A, Panteleakou Z, et al: Preferential expression of IGF-1Ec (MGF) transcript in cancerous tissues of human prostate: evidence for a novel and autonomous growth factor activity of MGF E peptide in human prostate cancer cells. Prostate. 70:1233–1242. 2010. View Article : Google Scholar
31	Philippou A, Armakolas A and Koutsilieris M: Evidence for the possible biological significance of the IGF-1 gene alternative splicing in prostate cancer. Front Endocrinol (Lausanne). 4:312013.PubMed/NCBI
32	Straub L: Beyond the transcripts: what controls protein variation? PLoS Biol. 9:e10011462011. View Article : Google Scholar : PubMed/NCBI
33	Vogel C and Marcotte EM: Insights into the regulation of protein abundance from proteomic and transcriptomic analyses. Nat Rev Genet. 13:227–232. 2012.PubMed/NCBI
34	Klevebring D, Fagerberg L, Lundberg E, Emanuelsson O, Uhlén M and Lundeberg J: Analysis of transcript and protein overlap in a human osteosarcoma cell line. BMC Genomics. 11:6842010. View Article : Google Scholar : PubMed/NCBI
35	Dengler MA, Weilbacher A, Gutekunst M, et al: Discrepant NOXA (PMAIP1) transcript and NOXA protein levels: a potential Achilles’ heel in mantle cell lymphoma. Cell Death Dis. 5:e10132014.PubMed/NCBI
36	Mole S, McFarlane M, Chuen-Im T, Milligan SG, Millan D and Graham SV: RNA splicing factors regulated by HPV16 during cervical tumour progression. J Pathol. 219:383–391. 2009. View Article : Google Scholar : PubMed/NCBI
37	Condemine W, Takahashi Y, Zhu J, et al: Characterization of endogenous human promyelocytic leukemia isoforms. Cancer Res. 66:6192–6198. 2006. View Article : Google Scholar : PubMed/NCBI
38	Nojima T, Oshiro-Ideue T, Nakanoya H, et al: Herpesvirus protein ICP27 switches PML isoform by altering mRNA splicing. Nucleic Acids Res. 37:6515–6527. 2009. View Article : Google Scholar : PubMed/NCBI
39	Liu Y, Tong Z, Li T, et al: Hepatitis B virus × protein stabilizes amplified in breast cancer 1 protein and cooperates with it to promote human hepatocellular carcinoma cell invasiveness. Hepatology. 56:1015–1024. 2012.
40	Kalamvoki M and Roizman B: HSV-1 degrades, stabilizes, requires, or is stung by STING depending on ICP0, the US3 protein kinase, and cell derivation. Proc Natl Acad Sci USA. 111:E611–E617. 2014. View Article : Google Scholar : PubMed/NCBI