PLK-1 Silencing in Bladder Cancer by siRNA Delivered With Exosomes

详细信息    查看全文

• 作者：Kristin A. Greco^a ; Carrie A. Franzen^a ; Kimberly E. Foreman^b ; ^c ; Robert C. Flanigan^a ; Paul C. Kuo^c ; ^d ; Gopal N. Gupta^a ; ^c ; ^d ; ^e ; ^{gogupta@lumc.edu" class="auth_mail" title="E-mail the corresponding author}
• 刊名：Urology
• 出版年：2016
• 出版时间：May 2016
• 年：2016
• 卷：91
• 期：Complete
• 页码：241.e1-241.e7
• 全文大小：1360 K

文摘

To use exosomes as a vector to deliver small interfering ribonucleic acid (siRNA) to silence the polo-like kinase 1 (PLK-1) gene in bladder cancer cells.

Materials and Methods

Exosomes were isolated from both human embryonic kidney 293 (HEK293) cell and mesenchymal stem cell (MSC) conditioned media. Fluorescently labeled exosomes were co-cultured with bladder cancer and normal epithelial cells and uptake was quantified by image cytometry. PLK-1 siRNA and negative control siRNA were loaded into HEK293 and MSC exosomes using electroporation. An invasive bladder cancer cell line (UMUC3) was co-cultured with the electroporated exosomes. Quantitative reverse transcriptase polymerase chain reaction was performed. Protein analysis was performed by Western blot. Annexin V staining and MTT assays were used to investigate effects on apoptosis and viability.

Results

Bladder cancer cell lines internalize an increased percentage of HEK293 exosomes when compared to normal bladder epithelial cells. Treatment of UMUC3 cells with exosomes electroporated with PLK-1 siRNA achieved successful knockdown of PLK-1 mRNA and protein when compared to cells treated with negative control exosomes.

Conclusion

HEK293 and MSC exosomes were effectively used as a delivery vector to transport PLK-1 siRNA to bladder cancer cells in vitro, resulting in selective gene silencing of PLK-1. The use of exosomes as a delivery vector for potential intravesical therapy is attractive.