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Q. Qiu; Inhibitory Effect of Antisense Oligonucleotides on EGFR Expression , Proliferation and Migration of Retinal Pigment Epithelium Cells In Vivo . Invest. Ophthalmol. Vis. Sci. 2003;44(13):2342.
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Purpose: To study the effect of antisense epidermal growth factor receptor(EGFR) oligonucleotides (ODN) transfection on EGFR expression, proliferation and migration of cultured retinal pigment epithelium(RPE) Methods: Using cationic liposome-mediated method, antisense EGFR ODN were introduced into RPE, EGFR gene and protein expression were detected by semi-quanitative RT-PCR and ELISA, respectively. Cell hyperplasia was evaluated by cell cycle analysis and MTT.By using an in vitro wound healing model in which a small area of a confluent monolayer of RPE cells was denuded, Migration was measured as the number of cells that had entered the denuded area. Results: The protein expression of EGFR was inhibited effectively at 24h by 66.45 (P0.01)Transfection of antisense EGFR ODN inhibited EGFR mRNA expression by 35.2. The cell migration was inhibited effectively by antisense oligodeoxynucleotides with lipofectin by 75.27%. The cell proliferation was inhibited effectively by antisense oligodeoxynucleotides with lipofectin by 40 %.Cell cycle analysis showed a significant decrease in S phase and a remarkable increase in G1/G0 fraction after exposure to antisense EGFR ODN in comparison with the cells with missense oligonucleotide treatment at the corresponding time. The missense oligodeoxynucleotides showed no such effect( P0.01 ). Conclusion: The transfection of antisense EGFR ODN into cultured RPE effectively suppressed EGFR mRNA and protein expression and inhibited the RPE proliferation and migration. Antisense oligodeoxynucleotides conjugated with lipofectin might be a hopeful method to inhibit the migration and proliferation of RPE by inhibiting the expression of EGFR. Key words: Antisense oligonucleotides;Epidermal growth factor receptor; Migration;Expression; Hyperplasia
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