Summary
Gene amplification and enhanced expression of the epidermal growth factor receptor (EGFR) represent the major molecular genetic alteration in glioblastomas and it may play an essential role in cell growth and in the carcinogenic process. On the other hand, the nuclear suppressor proteins PML and p53 are also known to play critical roles in cancer development and in suppressing cell growth. Here we report that, in glioblastoma cells with defective EGFR function, the expressions of both promyelocytic leukaemia (PML) and p53 were altered. Cells that were transfected with the antisense-cDNA of EGFR were found to have more cells in G1 and fewer cells in S phase. In addition, the transfected cells were found to be non-responsive to EGF-induced cell growth. Interestingly, the expression of the suppressors p53 and PML were found to be significantly increased by immunohistochemical assay in the antisense-EGFR cells. Moreover, the PML expression in many of the cells was converted from the nuclear dot pattern into fine-granulated staining pattern. In contrast, the expressions of other cell cycle regulated genes and proto-oncogene, including the cyclin-dependent kinase 4 (cdk4), retinoblastoma, p16INK4a and p21H-ras, were not altered. These data indicate that there are specific inductions of PML and p53 proteins which may account for the increase in G1 and growth arrest in antisense-EGFR treated cells. It also indicates that the EGF, p53 and PML transduction pathways were linked and they may constitute an integral part of an altered growth regulatory programme. The interactions and cross-talks of these critical molecules may be very important in regulating cell growth, differentiation and cellular response to treatment in glioblastomas.
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References
Bandyopadhyay, D, Maddal, M, Adam, L, Mendelsohn, J & Kumar, R (1998). Physical interaction between epidermal growth factor receptor and DNA-dependent protein kinase in mammalian cells. J Biol Chem 273: 1568–1573.
Canman, CE, Chen, CY, Lee, MH & Kastan, MB (1994). DNA damage responses: p53 induction, cell cycle perturbations and apoptosis. Cold Spring Harb Sym Quan Biol LIX: 277–286.
Cao, H, Lei, ZM, Bian, L & Rao, CV (1995). Functional nuclear epidermal growth factor receptors in human choriocarcinoma JEG-3 cells and normal human placenta. Endocrinology 136: 3163–3172.
Chakrabarty, S, Rajagopal, S & Huang, S (1995). Expression of antisense epidermal growth factor receptor RNA downregulates the malignant behavior of human colon cancer cells. Clin Exp Metastasis 13: 191–195.
Chan, JYH, Li, L, Fan, YH, Mu, ZM, Zhang, WW & Chang, KS (1997). Cell-cycle regulation of DNA damage-induced expression of the suppressor gene PML. Biochem Biophys Res Commun 240: 640–646.
Chan, JYH, Chin, W, Liew, CT, Chang, KS & Johnson, PJ (1998). Altered expression of the growth- and transformation-suppressor PML in human hepatocellular carcinomas and in hepatitis tissues. Eur J Cancer 34: 1015–1022.
Chang, KS, Fan, YH, Andreeff, M, Liu, J & Mu, ZM (1995). The PML gene encodes a phosphoprotein associated with the nuclear matrix. Blood 85: 3646–3653.
De Giovanni, C, Landuzzi, L, Frabetti, F, Nicoletti, G, Griffoni, C, Rossi, I, Mazzotti, M, Scotto, L, Nanni, P & Lollini, PL (1996). Antisense epidermal growth factor receptor transfection impairs the proliferative ability of human rhabdomyosarcoma cells. Cancer Res 56: 3898–3901.
Ded, SP, Munoz, RM, Brown, DR, Subler, MA & Sumitra, D (1994). Wild-type human p53 activates the human epidermal growth factor receptor promoter. Oncogene 9: 1341–1349.
Dixit, M, Yang, JL, Poirier, MC, Price, JO, Andrews, PA & Arteaga, CL (1997). Abrogation of cisplatin-induced programmed cell death in human breast cancer cells by epidermal growth factor antisense RNA. J Natl Cancer Inst 89: 365–373.
Everett, RD & Maul, GG (1994). HSV-1 IE protein Vmw110 causes redistribution of PML. EMBO J 13: 5062–5069.
Gomez-Manzano, C, Fueyo, J, Kyritsis, AP, McDonnell, TJ, Steck, PA, Levin, VA & Yung, WK (1997). Characterization of p53 and p21 functional interactions in glioma cells en route to apoptosis. J Natl Cancer Inst 89: 1036–1044.
Hartwell, LH & Kastan, MB (1994). Cell cycle control and cancer. Science 266: 1821–1828.
Koken, MHM, Linares-Cruz, G, Quignon, F, Viron, A, Chelbi-Alix, MK, Sobczak-Thepot, J, Juhlin, L, Degos, L, Calvo, F & de The, H (1995). The PML growth-suppressor has an altered expression in human oncogenesis. Oncogene 10: 1315–1324.
Kordek, R, Biernat, W, Alwasiak, ZJ, Maculewicz, R, Yanagihara, R & Liberski, P (1995). p53 protein and epidermal growth factor receptor expression in human astrocytomas. J Neuro-Oncol 26: 11–16.
Lamond, AI & Earnshaw, WC (1998). Structure and function in the nucleus. Science 280: 547–553.
Libermann, TA, Nusbaum, HR, Razon, N, Kris, R, Lax, I, Soreq, H, Whittle, N, Waterfield, MD, Ullrich, A & Schlessinger, J (1985). Amplification, enhanced expression and possible rearrangement of EGF receptor gene in primary human brain tumors of glial origin. Nature 313: 144–147.
Liu, JH, Mu, ZM & Chang, KS (1995). PML suppresses oncogenic transformation of NIH-3T3 cells by activated neu. J Exp Med 181: 1965–1973.
Liu, T, Chen, J & Zeng, C (1995). Effects of antisense epidermal growth factor and its receptor retroviral expression vectors on cell growth of human pancreatic carcinoma cell line. Chin Med J (Engl) 108: 653–659.
Louis, DN (1997). A molecular genetic model of astrocytoma. Brain Pathol 7: 755–764.
Ludes-Meyers, JH, Subler, MA, Shivakumar, CV, Munoz, RM, Jiang, P, Bigger, JE, Brown, DR, Deb, SP & Deb, S (1996). Transcriptional activation of the human epidermal growth factor receptor promoter by human p53. Mol Cell Biol 16: 6009–6019.
Maul, GG, Yu, E, Ishov, AM & Epstein, AL (1995). Nuclear domain 10 (ND10) associated proteins are also present in nuclear bodies and redistribute to hundreds of nuclear sites after stress. J Cell Biochem 59: 498–513.
Mendelsohn, J & Fan, Z (1997). Epidermal growth factor receptor family and chemosensitization. J Natl Cancer Inst 89: 341–343.
Montano, X (1997). p53 associates with trk tyrosine kinase. Oncogene 15: 245–256.
Morgenstern, JP & Land, H (1990). Advanced mammalian gene transfer: high titre retroviral vectors with multiple drug selection markers and a complementary helper-free packaging cell line. Nucleic Acid Res 18: 3587–3596.
Moroni, MC, Willingham, MC & Beguinot, L (1992). EGF-R antisense RNA blocks expression of the epidermal growth factor receptor and suppresses the transforming phenotype of a human carcinoma cell line. J Biol Chem 267: 2714–2722.
Mu, ZM, Chin, KV, Liu, JH, Lozano, G & Chang, KS (1994). PML, a growth suppressor disrupted in acute promyelocytic leukemia. Mol Cell Biol 14: 6858–6867.
Nagane, M, Coufal, F, Lin, H, Bogler, O, Cavenee, WK & Huang, HJS (1996). A common mutant epidermal growth factor receptor confers enhanced tumorigenicity on human glioblastoma cells by increasing proliferation and reducing apoptosis. Cancer Res 56: 5079–5086.
Ng, HK & Lam, PYP (1998). The molecular genetics of central nervous system tumours. Pathology 30: 196–202.
Puvion-Dutilleul, F, Chelbi-Alix, M, Koken, M, Quignon, F, Puvion, E & de The, H (1995). Adenovirus infection induces rearrangements in the intranuclear distribution of the nuclear body-associated PML protein. Exp Cell Res 218: 9–16.
Schober, R, Bilzer, T, Waha, A, Reifenberger, G, Wechsler, W, von Deimling, A, Wiestler, OD, Westphal, M, Kemshead, JT & Vega, F (1995). The epidermal growth factor receptor in glioblastoma: genomic amplification, protein expression, and patient survival data in a therapeutic trial. Clin Neuropathol 14: 169–174.
Sidransky, D & Hollstein, M (1996). Clinical implications of the p53 gene. Annu Rev Med 47: 285–301.
Smith, ML & Fornace, AJ (1995). Genomic instability and the role of p53 mutations in cancer cells. Curr Opin Oncol 7: 69–75.
Tang, P, Steck, PA & Yung, WK (1997). The autocrine loop of TGF-alpha/EGFR and brain tumors. J Neuro-Oncol 35: 303–314.
Terris, B, Baldin, V, Dubios, S, Degott, C, Flejou, JF, Henin, D & Dejean, A (1995). PML nuclear bodies are general targets for inflammation and cell proliferation. Cancer Res 55: 1590–1597.
Tian, XX, Lam, PYP, Chan, J, Pang, JCS, To, SST, Di-Tomaso, E & Ng, HK (1998). Antisense EGF receptor RNA transfection in human malignant glioma cells leads to inhibition of proliferation and induction of differentiation. Neuropath Appl Neurobiol 24: 389–396.
Vallian, S, Gaken, JA, Trayner, ID, Gingold, EB, Kouzarides, T, Chang, KS & Farzaneh, F (1997). Transcriptional repression by the promyelocytic leukemia protein PML. Exp Cell Res 237: 371–382.
Wang, S, Lee, RJ, Cauchon, G, Gorenstein, DG & Low, PS (1995). Delivery of antisense oligodeoxyribonucleotides against the human epidermal growth factor receptor into cultured KB cells with liposomes conjugated to folate via polyethylene glycol. Proc Natl Acad Sci USA 92: 3318–3322.
Wong, AJ, Bigner, SH, Kinzler, KW, Hamilton, SR & Vogelstein, B (1987). Increased expression of the EGF receptor gene in malignant gliomas is invariably associated with gene amplification. Proc Natl Acad Sci USA 84: 6899–6903.
Yarden, Y & Ullrich, A (1988). Growth factor receptor tyrosine kinases. Annu Rev Biochem 57: 443–478.
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Tian, X., Chan, J., Pang, J. et al. Altered expression of the suppressors PML and p53 in glioblastoma cells with the antisense-EGF-receptor. Br J Cancer 81, 994–1001 (1999). https://doi.org/10.1038/sj.bjc.6690798
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DOI: https://doi.org/10.1038/sj.bjc.6690798
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