title: Combined EGFR and c-Src antisense oligodeoxynucleotides encapsulated with PAMAM Denderimers inhibit HT-29 colon cancer cell proliferation.
abstract: Colon cancer continues to be one of the most common cancers, and the importance and necessity of new therapies needs to be stressed. The most important proto-oncogen factors for colon cancer appear to be epidermal growth factor receptor, EGFR, and c-Src with high expression and activity leading to tumor growth and ultimately to colon cancer progression. Application of c-Src and EGFR antisense agents simultaneously should theoretically therefore have major benefit. In the present study, anti-EGFR and c-Src specific antisense oligodeoxynucleotides were combined in a formulation using PAMAM dendrimers as a carrier. Nano drug entry into cells was confirmed by flow cytometry and fluorescence microscopy imaging and real time PCR showed gene expression of c-Src and EGFR, as well as downstream STAT5 and MAPK-1 with the tumor suppressor gene P53 to all be downregulated. EGFR and c-Src protein expression was also reduced when assessed by western blotting techniques. The effect of the antisense oligonucleotide on HT29 cell proliferation was determined by MTT assay, reduction beijng observed after 48 hours. In summary, nano-drug, anti-EGFR and c-Src specific antisense oligodeoxynucleotides were effectively transferred into HT-29 cells and inhibited gene expression in target cells. Based on the results of this study it appears that the use of antisense EGFR and c-Src simultaneously might have a significant effect on colon cancer growth by down regulation of EGFR and its downstream genes.
authors.affiliation: Department of Biochemistry, Faculty of Medicine, Kerman University of Medical Sciences, Kerman, Tabriz, Iran.
authors.forename: Ali Reza
authors.initials: AR
authors.lastname: Nourazarian
authors.forename: Ahmad Gholamhoseinian
authors.initials: AG
authors.lastname: Najar
authors.forename: Safar
authors.initials: S
authors.lastname: Farajnia
authors.forename: Ahmad Yari
authors.initials: AY
authors.lastname: Khosroushahi
authors.forename: Roghiyeh
authors.initials: R
authors.lastname: Pashaei-Asl
authors.forename: Yadollah
authors.initials: Y
authors.lastname: Omidi
mesh_terms: D000704:Analysis of Variance; D049109:Cell Proliferation / Q000187:drug effects*; D050091:Dendrimers; D015536:Down-Regulation / Q000187:drug effects; D066246:ErbB Receptors / Q000235:genetics* / Q000378:metabolism; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects*; D019073:HT29 Cells; D006801:Humans; D019950:Mitogen-Activated Protein Kinase 1 / Q000235:genetics; D016376:Oligonucleotides, Antisense / Q000235:genetics / Q000494:pharmacology*; D050799:STAT5 Transcription Factor / Q000235:genetics; D016159:Tumor Suppressor Protein p53 / Q000235:genetics; D019061:src-Family Kinases / Q000235:genetics* / Q000378:metabolism
chemical_list: D050091:Dendrimers; D016376:Oligonucleotides, Antisense; C104700:PAMAM Starburst; D050799:STAT5 Transcription Factor; D016159:Tumor Suppressor Protein p53; D066246:ErbB Receptors; D019061:src-Family Kinases; C535150:MAPK1 protein, human; D019950:Mitogen-Activated Protein Kinase 1