Análisis de la expresión del gen IGF-I identificado como oncogen en células de glioma de ratón parentales y transfectadas usando estrategia de terapia génica por triple hélice y antisentido

Abstract

Cancer is a worldwide public health problem due to its mortality rate. It was shown that in Colombia we have 2-19 glioblastoma patients / 100,000 inhabitants per year. Glioblastoma constitutes a primary glial malignant tumor of CNS; the research investigations of glioblastoma have demonstrated a significant increase in IGF-I gene expression in this brain tumor. The objective of this study was to use gene therapy technology of either antisense, AS, or triple helix, TH, type to explore the levels of IGF-I gene expression in somatic transfected cells. The cells of rat C6 glioma line were transfected in vitro with IGF-I AS, IGF-I TH expression vectors, and with control pMT-EP empty vector, using lipofectin reagent. The IGF-I gene expression in cultured cells was investigated by RT-PCR performed with TaqMan probes; a 800pb fragment of the promoter region of this gene was evaluated by capillary electrophoresis. A decrease in IGF-I gene expression, as 80% and 60%, was demonstrated in case of used TH and AS technologies, respectively. The sequences of 800 bp fragment of IGF-I gene studied in human normal samples, front brain tumor samples and glioma samples, had no demonstrated the mutations. These promising findings allow to investigate the human cells in vitro in a new way. The presented in vitro results can be applied for preparation of cell vaccines to be used in vivo in cell gene therapy.