Design of antisense oligonucleotides against twist1 gene and evaluation of their anti-invasive effects in prostate cancer cell lines

Abstract

Introduction: Prostate cancer is one of the most common cancers and the second major cause of mortality in men. Different researches have shown that the overexpression of a gene called twist1 leads to initiation of metastasis process in this cancer. TWIST1 protein triggers this process through stimulating the transition pathway of cells from epithelial to mesenchymal tissue.Materials and Methods: In this study, four oligonucleotides of antisense RNA have been designed for twist1 gene, and its anti-metastatic effect was examined in two cell lines PC3 and LNCaP. The antisense oligonucleotides (ASOs) were designed as single strands with a length of 20 nucleotides and chosen from ASOs suggested by Soligo program. The ASOs were synthesized in phosphorothioated form. MTT assay was used for evaluating the ASOs cytotoxicity effect on PC3 and LNCaP cell lines. The cell lines were transfected with 500 nmol of antisense oligonucleotides using cationic polymer turbofect and incubated for 48 hours, and then, their invasive ability were measured by CytoSelect™ Cell Invasion Assay Kit.Results: The anti-invasive effect of ASOs in LNCaP and PC3 had a significant difference. This effect was more significant in LNCaP cell line as compared to PC3. The most anti- invasive effect was observed in LNCaP cell line (50%).Conclusion: According to the results, Antisense oligonucleotides were effective in decreasing the invasion ability of two cell lines PC3 and LNCaP and therfore can be considered as a good candidate for preventing the prostate cancer metastasis.