Targeting Melanoma Growth and Metastasis with Systemic Delivery of Liposome-Incorporated Protease-Activated Receptor-1 Small Interfering RNA

Department of Cancer Biology, The University of Texas MD Anderson Cancer Center, Houston, Texas 77030, USA.
Cancer Research (Impact Factor: 9.33). 11/2008; 68(21):9078-86. DOI: 10.1158/0008-5472.CAN-08-2397
Source: PubMed


The thrombin receptor [protease-activated receptor-1 (PAR-1)] is overexpressed in highly metastatic melanoma cell lines and in patients with metastatic lesions. Activation of PAR-1 leads to cell signaling and up-regulation of genes involved in adhesion, invasion, and angiogenesis. Herein, we stably silence PAR-1 through the use of lentiviral short hairpin RNA and found significant decreases in both tumor growth (P < 0.01) and metastasis (P < 0.001) of highly metastatic melanoma cell lines in vivo. The use of viruses for therapy is not ideal as it can induce toxic immune responses and possible gene alterations following viral integration. Therefore, we also used systemic delivery of PAR-1 small interfering RNA (siRNA) incorporated into neutral liposomes [1,2-dioleoyl-sn-glycero-3-phosphatidylcholine (DOPC)] to decrease melanoma growth and metastasis in vivo. Significant decreases in tumor growth, weight, and metastatic lung colonies (P < 0.001 for all) were found in mice treated with PAR-1 siRNA-DOPC. The in vivo effects of PAR-1 on invasion and angiogenesis were analyzed via immunohistochemistry. Concomitant decreases in vascular endothelial growth factor, interleukin-8, and matrix metalloproteinase-2 expression levels, as well as decreased blood vessel density (CD31), were found in tumor samples from PAR-1 siRNA-treated mice, suggesting that PAR-1 is a regulator of melanoma cell growth and metastasis by affecting angiogenic and invasive factors. We propose that siRNA incorporated into DOPC nanoparticles could be delivered systemically and used as a new modality for melanoma treatment.

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    BioMed Research International 07/2014; 2014(1):895986. DOI:10.1155/2014/895986 · 2.71 Impact Factor
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    • "For western blot analyses, both hippocampi of all the mice from each of the experimental groups were obtained. Protein extraction, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and western blot were performed based on the manufacturer's instructions10,21). Equivalent amounts of protein were separated using 8% SDS-PAGE and transferred to PVDF membranes, followed by immunoblotting with the primary antibody. "
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    • "Although the results suggest that N-cadherin positively affects the regulation of the cell cycle and proliferation through activation of the AKT kinase pathway, further investigations are needed to describe the mechanism. Similarly, Villares et al., upon the observation that thrombin receptor (or protease-activated receptor-1, PAR-1) is overexpressed in highly metastatic melanoma cell lines, has evaluated the therapeutic potential of siRNA against PAR-1 [115]. The authors have observed a significant reduction of in vivo tumor growth as well as in the number of metastatic lung colonies. "
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