International Journal Of Bilogical Sciences
.: Home > International Journal Of Bilogical Sciences > 2015 > Volume 9 Number 5 > Xiao-yan Liu1, Qiu-su Tang2, Hong-chao Chen1, Xiao-ling Jiang1, Hong Fang1✉
Lentiviral miR30-based RNA Interference against Heparanase Suppresses Melanoma Metastasis with Lower Liver and Lung Toxicity
Xiao-yan Liu1, Qiu-su Tang2, Hong-chao Chen1, Xiao-ling Jiang1, Hong Fang1✉
1. Department of Dermatology, the First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, Zhejiang 310003, China 2. Department of Pathology, the First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, Zhejiang 310003, China
Aim: To construct short hairpin RNAs (shRNAs) and miR30-based shRNAs against heparanase (HPSE) to compare their safety and their effects on HPSE down-modulation in vitro and in vivo to develop a more ideal therapeutic RNA interference (RNAi) vector targeting HPSE.
Methods: First, we constructed shRNAs and miR30-based shRNAs against HPSE (HPSE-shRNAs and HPSE-miRNAs) and packed them into lentiviral vectors. Next, we observed the effects of the shRNAs on knockdown for HPSE expression, adhesion, migration and invasion abilities in human malignant melanoma A375 cells in vitro. Furthermore, we compared the effects of the shRNAs on melanoma growth, metastasis and safety in xenograft models.
Results: Our data showed that these artificial miRNAs targeting HPSE could be effective RNAi agents mediated by Pol II promoters in vitro and in vivo, although these miRNAs were not more potent than the HPSE-shRNAs. It was noted that obvious lung injuries, rarely revealed previously, as well as hepatotoxicity could be caused by lentivirus-mediated shRNAs (LV shRNAs) rather than lentivirus-mediated miRNAs (LV miRNAs) in vivo. Furthermore, enhanced expression of pro-inflammatory cytokines IL-6 and TGF-β1 and endogenous mmu-miR-21a-5p were detected in lung tissues of shRNAs groups, whereas the expression of mmu-let-7a-5p, mmu-let-7b-5p and mmu-let-7c-5p were down-regulated.
Conclusion: These findings suggest that artificial miRNAs display an improved safety profile of lowered lung injury or hepatotoxicity relative to shRNAs in vivo. The mechanism of lung injuries caused by shRNAs may be correlated with changes of endogenous miRNAs in the lung. Our data here increase the flexibility of a miRNA-based RNAi system for functional genomic and gene therapy applications.
RNA interference, microRNA(miRNA), heparanase, metastasis, safety
Date Deposited : 11 Apr 2015 10:02
Official URL: http://www.ijbs.com/ms/archive
Last Modified : 11 Apr 2015 10:02
Volume 9, Number 5, - 2015 , ISSN 1449-2288
Full Text Original